https://www.neosynbio.com/home daily 1.0 2023-03-07 https://images.squarespace-cdn.com/content/v1/57d6c55a2994cab476e9fb40/1588549806362-31G8OAKLZ692DUDF1WLJ/ca0012.jpg Home https://images.squarespace-cdn.com/content/v1/57d6c55a2994cab476e9fb40/1588546032089-BAULXE3CYR01AZBG9AAX/2015-08-12_home-goods-0052-v2-FINAL.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258161213-QQFT47P69746HM39NDEM/82253695_2771154009609526_4181759504334979072_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258557562-W7SAO7Q8J44RTD4GY3SB/36327649_1805867702804833_562509719273472000_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258225514-7N0SR5NGMDEPY8KZYN61/60561160_2289355657789366_5721832483444490240_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592188183812-FCZY901ETJLMM2V2X615/69484976_950266078640219_1643356677606473728_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592183045124-74OJL121RXOYOF80HNCZ/IMG_20200401_130616.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258603608-TOOSEX6BA8RF2NK67PBQ/87480863_2858195094238750_5711241875776602112_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258635448-MVZWOGW6ZXXCW3Z2TLUN/66037471_2378068962251368_5690479595845844992_o.jpg Home https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605836800415-7M6NG2JU5ZEGELGIUTI0/maxresdefault.jpg Home Igemers of the World Podcast https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592185979010-RSSKIKDACSZRLKU9LBIB/5dolla.PNG Home “DIY biologists from around the world hope their research will drive down the cost of insulin.” https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592186716418-NA4GK5YX6D7ODN99B9KW/publication.PNG Home The Open Insulin Project: A Case Study for ‘Biohacked’ Medicines https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592187406147-REMLGASP7EJN0COGIRZ4/image-asset.jpeg Home https://www.neosynbio.com/contact daily 0.75 2020-11-08 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604622965195-CVVJZ00E7KM65PNVJERF/image.jpg Consulting Private Industry & Investors/ Government https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604622930333-3XR16F9RQ94ZN8IFZ4LR/image.jpg Consulting Open Source/ Not-For-Profit Pro-Bono https://www.neosynbio.com/protocols daily 0.75 2020-11-27 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597800127661-YDDHKB1ZO8992YIECWIJ/image-asset.jpeg All Protocols - The Sterile Workspace https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597729771693-AV9DIQUF3TXY6OOLBPAO/T--Sydney_Australia--Results1.jpg All Protocols - HOST SELECTION CRITERIA https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597730987656-D3KC8ZCDGS973HXVLZRK/insulin.jpg All Protocols - CULTURING cONDITIONS https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597731046282-M4GQTBLM036D4NIJ997X/HAND.jpg All Protocols - mIXING MEDIA & POURING PLATES https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597786707463-EN3DC5R601USY6B2WEMR/is-it-safe-to-use-Bunsen-burner-in-a-biological-safety-cabinet.jpg All Protocols - sTERILE tECHNIQUE https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597787306611-MPNF6FUG3ZQH8BPHAYEA/IMG_20191216_120843.jpg All Protocols - Buffer Mixing https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603062012273-8VDCQ3LQMZ3OCPRXIP1O/Dna3.jpg All Protocols - DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797043131-OLUKDBQXRM4NEAVG5669/IMG_20190409_120627.jpg All Protocols - Purification of Genomic DNA https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597796835006-5HWICXOQTQTGJZC9WYOA/Backbone.jpg All Protocols - plasmids101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797322389-RFH17Z0GJOUGLETMJ2Z4/49825983_2093085314083069_6283794027452039168_o.jpg All Protocols - purification of plasmid DNA (Miniprep aka. Ethanol Lysis) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797147471-G18C9A9M5J544Y0URJK1/IMG_20190905_140727.jpg All Protocols - PlasmiD Insertion https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797211093-76P4B3SCSYFUIT6KX8OM/49175166_2081528401905427_7542656853239398400_o.jpg All Protocols - aGAROSE geL eLECTROPHORESIS https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797652320-6F39AHF5058MDTD7RA3V/sebgel.jpg All Protocols - gEL iLLUMINATION & PHOTOGRAPHY https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597797620069-HS7802PW2NU6SUJ1DP9S/Gelfun.jpg All Protocols - Restriction Enzyme Digest of Plasmid & Insert DNA https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597798296036-Q2AQAKAPFK520E6G3DNW/70928786_2530275270364069_7666269843129106432_o.jpg All Protocols - Ligation & Troubleshooting https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597798404578-COO7VJDPKTZBJURJ89LN/MVIMG_20190809_091538.jpg All Protocols - Polymerase Chain Reaction (PCR) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597798534098-DXKMPFQY64OC4Q7JRSDY/3%2527clamp.jpg All Protocols - Primer Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597798726828-U02V50OIZX7KH8QUOMUZ/MVIMG_20190812_115408.jpg All Protocols - Gradient PCR https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597799096659-46FIZNZYZ5230L3RV7IK/Insulingel.jpg All Protocols - Confirmation of Edits https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597799240819-26RM5EFR92NL8L2IZ1BM/Flankingaddition.jpg All Protocols - flanking sequence addition https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606440028227-W1ZI8XL01X4D57WC5P8P/Gibson%2BAssembly.jpg All Protocols - Gibson Assembly https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598845365549-7Q3G3XF3K5WM6BP8YHRI/image-asset.jpeg All Protocols - Golden Gate Assembly https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598845638244-B02SZMZMNXSE25WBWKRQ/32207602_1750046595053611_7302529058535899136_o.jpg All Protocols - Site Directed Mutagenesis https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598846472868-II770HH2NVBFFWX8CY50/Designbuildtestlearn.jpg All Protocols - Gblock Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598845896204-6NFJCH3LX5N9LQ1UPZ2M/Backbone.jpg All Protocols - Backbone Selection https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598845742783-L63FU1PH6M8GSV1Z4RW0/ADN_Cas9_Feldan.jpg All Protocols - CRISPR/CAS9 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597888291437-IGTAKKGIJEOO7DPSIDGF/51265829_2124280340963566_5495482364005974016_n.jpg All Protocols - Proteins101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597888233832-YLN7Z2NYVRQWR9SFL3BS/82253695_2771154009609526_4181759504334979072_o.jpg All Protocols - Induction of Protein Expression https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597893733326-EQFVAQKXHMVSEDAXT5S1/beads.jpg All Protocols - Cell Lysis https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597897639619-2EPETAE9OQ07UQ53RELR/F90154_p.eps-650.jpg All Protocols - Affinity Tag Purification https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597888539554-OVUMGKYSV5Y6LDOJ4BDF/T--Sydney_Australia--wk26SDSpage.jpg All Protocols - SDS-PAGE https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606435448521-HSL97I7PP1SPSBLOIJD1/Coming%2BSoon.jpg All Protocols - ELISA https://www.neosynbio.com/projects daily 0.75 2026-02-02 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598492699584-UQ2W4FN8LDZKZPC3DONA/image-asset.jpeg All Projects - The Australian team of ThE Open Insulin Project https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598492910227-ERWDHPXBIOMD6ELRBW1W/82253695_2771154009609526_4181759504334979072_o.jpg All Projects - Welcome to development hell… CRISPR/CAS9 Demonstration Kit https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598493167472-J1DTDYKJ6X1SGFFP49JV/36327649_1805867702804833_562509719273472000_o.jpg All Projects - Using plastic until I develop a cell printer 3D Printed Prosthetic Limbs https://www.neosynbio.com/plastic-circular-economy daily 0.75 2020-11-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598406108036-5AOO0V3D9GBBU432ZGNV/3c757c9e7ad496ee92590b00ac0cd2c6559e8281.jpg Plastic Circular Economy Image credit: https://www.smh.com.au/national/this-is-d-day-for-the-nation-s-recycling-crisis-20191031-p5365d.html https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598412016119-2T8N35IKKLSBHKZ9EEGX/PET_polymer20.png Plastic Circular Economy https://ambermd.org/tutorials/advanced/tutorial27/pet.htm https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598413480227-6NHSSGJLOIHTWTHK9P3B/plastic+by+type.PNG Plastic Circular Economy Image and Statistics credit: https://www.abs.gov.au/ausstats/abs@.nsf/mf/4602.0.55.005#:~:text=Australian%20households%20generated%2045.6%25%20(1.2,%2C%20267%2C925%20tonnes%20(22.9%25). https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598833143265-36I6K6ETVUWH5CPHN5AK/Cumate+Inducible+Expression+PET+Depolymerase+F243I+D238C+S283C+Map.png Plastic Circular Economy https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598835306909-O1IKL0W29VMSSKVTWBTB/PET+monomers.jpg Plastic Circular Economy Image credit: https://chem.libretexts.org/Courses/Brevard_College/CHE_202%3A_Organic_Chemistry_II/02%3A_Carboxylic_Acids_and_Esters/2.07%3A_Preparation_of_Esters https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598405772806-PCR5DBO7GXGTJFEVT3CF/98176752_3031830970208494_7369318900955086848_n+%281%29.png Plastic Circular Economy https://www.neosynbio.com/equipment daily 0.75 2020-12-07 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600300689529-63R8B491LMX6C2I6UMUO/image-asset.jpeg Equipment - Gloves https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600300902659-XO24HUJK60O6SVO5UZEM/5dolla.jpg Equipment - Lab Coat https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600300983632-ORPISGYJSWE0R3YPGQWT/image-asset.jpeg Equipment - Goggles https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600295532632-IT40S57V6MZ2E45ARS4J/is-it-safe-to-use-Bunsen-burner-in-a-biological-safety-cabinet.jpg Equipment - inoculation Wand https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600295699570-RBIGHX7DJZIOC2YFAEWK/69484976_950266078640219_1643356677606473728_o.jpg Equipment - Micropipettes https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600295869811-LVC6MZHNZNCDML0N955P/is-it-safe-to-use-Bunsen-burner-in-a-biological-safety-cabinet.jpg Equipment - Bunsen Burner https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606178739555-VM8S5EDT4JX2KXOLYTIY/image.jpg Equipment - Sundry https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606178845183-CRWL6JLIG3065P34RN9W/laminar%2Bflow%2Bhood.jpg Equipment - Laminar Flow hood https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606178957149-NYH7BGVCG8KQM2M5ZX86/biosafety%2Bcabinet.jpg Equipment - Biosafety Cabinet https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601341337929-10G3NHVHH4AXLMYWNH6Y/0-02-07-a186682c71fe6ac1af68bba287f915b50f603928f587c4f14aba81feb2359289_eaba7a34.jpg Equipment - Weighing Scale https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601341590687-7UBNGYJ9RMUXOAMHEG2J/image-asset.jpeg Equipment - Glassware https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601341636720-VVFHKNWU2DZ5I5BOU50Q/Pipeboy-acu-2-Showcase.jpg Equipment - Pipetteboi https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605834524476-RTGI2E72L1RPDR497TX9/0005876_cell-spreaders-glass.jpg Equipment - Glass Spreader https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605834586525-33MONRJKF1QPGBZGWLWA/093fbe7ec374f8703be9841130bef486b845b30c.jpg Equipment - Vortex https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605834751610-7IL4MAA1OZ2T47LOA1L4/CX-ims-100-xopen-400x400.jpg Equipment - Ice Machine & Box https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301104020-RWU7O2AOTVYD9XTAFUKE/57191754_2242092495849016_3398564821975695360_o.jpg Equipment - RO/MILLIQ Water https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301284601-RCAW2E9W613ZRJQIZETN/79924367_2702259549832306_7117383652744364032_o.jpg Equipment - pH Meter https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301350005-8DB4LIAYOPT2QADE2SK1/vials-1781316_960_720.jpg Equipment - Liquid Waste https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600295985171-YM4RV7H9JWWP0688Y2PJ/image-asset.jpeg Equipment - Disinfectant SPray https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600296087364-5DHUBIQH32SV3V6R9VE8/IMG_20190905_140727.jpg Equipment - Petri Dishes https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600296163582-AG02RQGWORFCHHJ8K3JF/82253695_2771154009609526_4181759504334979072_o.jpg Equipment - Tubes & Tube Racks https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600296407105-CKOL6C88MZ8GTI0VHGV1/IMG_20200513_102728.jpg Equipment - Autoclave https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600296516229-XENHE094ZCSME5QCNBGI/70928786_2530275270364069_7666269843129106432_o.jpg Equipment - Fridge & fREEZERS https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600296864159-KONVOWINON27YXU3EWH8/MVIMG_20190409_102004.jpg Equipment - incubator https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600297165008-FROCVYV0J30QC7FDLP20/IMG_20190409_120627.jpg Equipment - Heat Block https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600297256959-W6ZA3XH1I8PZN611I2AG/growgrow.jpg Equipment - Hot Water Bath https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600297397211-9O6E3I67E7AILGU573B7/image-asset.jpeg Equipment - Microwave https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301409743-6TUMU61ZE85F2X63RD7D/centrifuge-3291253_1920.jpg Equipment - Centrifuge https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301568648-PROFRZ6S090SBM6SDO84/49825983_2093085314083069_6283794027452039168_o.jpg Equipment - Silica Spin Columns https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301502784-SNM1R2PC9LVDGIPJT7YQ/MVIMG_20190809_091538.jpg Equipment - ThermoCycler https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606088358087-N446LAL8RXQY07P1MLF4/Ovens-and-Furnaces-%2528parts%2529.jpg Equipment - Oven https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606088093656-ZBWL8FLOI841GS80RU4Q/gb-onda-spectrophotometer1-web.jpg Equipment - Spectrophotometer https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606087856828-4HHUMGFGXF7J8W4COJKB/ND2000008-800.jpg Equipment - Nanodrop/Qubit https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301688987-EANS2GVRGM47MBBAAJ3X/49175166_2081528401905427_7542656853239398400_o.jpg Equipment - DC Power Supply https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301790585-GVRR2OF1Z42D1AKSVGC9/33921060_1769725416419062_7881852548276027392_o.jpg Equipment - Gel Tray & Bath https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600301984983-A9605WYUDAKBIG0AA9XO/forwebsite.jpg Equipment - Illuminator https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600302157955-LOAWDC26AWFGDJAF7X34/IMG_20191101_094852.jpg Equipment - Imaging Enclosure https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600302313709-8VS92T1LS0K5B22L5XAU/beads.jpg Equipment - Bead Beater https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600302428612-L4UPJI9AXXF9L8YT8DLD/T--Sydney_Australia--wk26SDSpage.jpg Equipment - SDS-PAGE Enclosure https://www.neosynbio.com/open-insulin-australia daily 0.75 2021-08-17 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593735253244-AP9RIVY3ZTBQUB1E8AKF/insulin-syringe-1972843_1920.jpg Open Insulin Australia - Alex Kelly Open Insulin australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593737967539-NRH13OIEPW0LLB215FN2/test_vial_price.001.jpeg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593737968794-SM00VEU6ZX40RXOUDZSY/EMfcuAHXkAIPZ5u.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738226281-0QDOSCXDBU8LOEURPAL2/23592060_193046327925754_8312462357427357046_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738054210-2LUR2UEUNOFM29N2KVXW/DISCO.PNG Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738218982-IJXTC5G8LC65GZIL688A/19467917_146489222581465_632730073322427084_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738222066-G4PCC6W1S53Y02XFNDJ4/20449088_159578827939171_4443545517730807381_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738221278-36V5UVS7KZ2M44PF3XTT/20989000_166785417218512_7791198595918154144_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738222594-9PMFMAOSXT1EX3GH2A45/21167166_168719007025153_4034730261579523725_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738222853-N5N09UMDZVTYDVP5JZ05/22406217_182465598983827_1228240899760098323_n.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1593738223112-UJXIU5LO3OVQCUFPXGNQ/22365422_182465615650492_9022237063417834547_n.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259210312-TNGUEFA4QDO19X5T5DRR/30167559_1720519601339644_418541686919896391_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259205360-F5NX64B138MOQ4REQASH/29872714_1720653111326293_3962288052971227280_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259207684-46C70FT460DKE1FM24KB/29872650_1721371944587743_1092111695274620513_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259209776-4EVY7FWBEP6UL9JRBMF7/30171218_1721496734575264_5342347648564381590_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259554416-FY6LT2NT6M10MVW559JU/75486085_2624768120914783_4983679376521428992_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259557941-6M67GSE30OMCJCC9X9XZ/66037471_2378068962251368_5690479595845844992_o.jpg Open Insulin Australia https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594259595919-5VEG4HBNBHGHCP38Q27L/5dolla.PNG Open Insulin Australia https://www.neosynbio.com/crisprcas9-demonstration-kit daily 0.75 2020-07-10 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594258959170-O68VWM9X0WEZODEGE0PJ/30051933_1728174887240782_3367079503584516366_o.jpg CRISPR/CAS9 Demonstration Kit Same cells, same plasmid. One single letter of code changed to induce the colour change. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594340141316-ZH1VU5JZALAPJRVNHLQM/Schematic-representation-of-CRISPR-Cas9-mediated-genome-editing-a-Schematic-of-CRISPR.png CRISPR/CAS9 Demonstration Kit https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594340821624-JU954L2SXT1IQ86WWLNC/CRISPR_CAS9+2.0+-+Superfolded+GFP+%281%29.jpg CRISPR/CAS9 Demonstration Kit Perhaps there were early hints that things weren’t going to be easy… https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594343146902-K4SPZ603PFXGJIQ6B6QG/protospacer.png CRISPR/CAS9 Demonstration Kit The lowercase t is the change induced by the Site-Directed Mutagenesis. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594343673081-IA7BLJCTRHFMFW8WWJM7/30051933_1728174887240782_3367079503584516366_o.jpg CRISPR/CAS9 Demonstration Kit https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1594342353253-82HD7YHYDAJPTZK1QD8X/Crispr+kit.png CRISPR/CAS9 Demonstration Kit At this point, the complexity of the planned system is starting to strain credulity… https://www.neosynbio.com/the-graduates-guide-to-synthetic-biology daily 0.75 2023-09-07 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600035179938-X7O0BRSM476WDH4FJHJA/two-hands-painting-the-planet-earth_BsB6xZcIDg.jpg The Graduate's Guide to Synthetic Biology The Graduate’s Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605661253907-F4NHHWK4CF98QOHPWOFI/Graduate%27s+Synbio+Guide+Layout+%282%29.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606247361448-R725JXNI4NSPAG8A3YSL/Bunsen.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1592266153078-Z03X73QU3GJCYJN9ZMST/image-asset.png The Graduate's Guide to Synthetic Biology Penicillin – the happiest accident of all? https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600045147364-215XVB4LEGOB3YQ6BBCJ/IMG_20190409_120627.jpg The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600045332251-2HO6MFKM947LMV366UI7/IMG_20191004_112639%2B%25281%2529.jpg The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600127875017-ST8A1ZWDP6K23F5KZKWH/92137009_2930683080323284_3175143582470242304_o.jpg The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600056947093-ELNYMBMQFTUDLC5M77W0/29594379_1706828996042038_1304135844277829514_n.jpg The Graduate's Guide to Synthetic Biology Image captured on a Biorad transilluminator. One of the first gels I imaged on my own, using what was once top-notch equipment. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600057420266-AIZTLENESEYNC8EJF0EQ/66037471_2378068962251368_5690479595845844992_o.jpg The Graduate's Guide to Synthetic Biology Image of gel taken using a Samsung S8+ smartphone camera using 3D printed light enclosure and Orange filter over a cracked, second-hand UV light box. Despite using my own phone and a jerry-rigged setup, the quality of the image is far better than those I was able to take in the commercial machine. Also far less fiddling around with corporate software. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600129469011-UUE1TOUMIGK6FGTVYSJY/pET-15b+Map1.png The Graduate's Guide to Synthetic Biology (click image for sequence download) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600129356477-W5388PTLRMXUYUU11AIL/enzymeset.PNG The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600129743636-XPF889W9ANJKJ7T2B7LC/pET-15b+Map2.png The Graduate's Guide to Synthetic Biology (click image for sequence download) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600130370012-PLE4ZDRTGSTTO19EV8EI/image-asset.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600130997376-XO8C7S7UERS0ETPUZLJZ/Insert+Fragment.PNG The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600131007340-4OTG5QEKU33FY7N6LZIM/Cloning.PNG The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600131235634-0WX9KJP6XCO29RTBZPV1/clean+ligation.PNG The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600131311702-49OOQ4L4NRCKJ54I1RAV/CPro-pET15b+Map.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600137629048-KZB8QKX4R0KITN8OBMI4/70928786_2530275270364069_7666269843129106432_o.jpg The Graduate's Guide to Synthetic Biology This is how I organise my lab freezer to help prevent novice hands from spoiling my expensive enzymes. An enzyme freezer MUST NOT have a defrost cycle. The easiest way to check if your freezer is appropriate is to check for large amounts of ice buildup. Lots of ice = probably doesn’t have a defrost cycle. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607381339993-TXCH2WX6AX03CMDGXJQA/Polymerase+Chain+Reaction+Cycle.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600145866076-OJ86YP3F0U7ISX11IQJ7/T--Sydney_Australia--InterlabcolonyPCR2.jpg The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600145968538-CYGNM20VAPMBGUP4N17Y/T--Sydney_Australia--Results3.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600206824655-TECY7DIF5WDGMO824D0I/1280px-Historic_cost_of_sequencing_a_human_genome.svg.png The Graduate's Guide to Synthetic Biology https://en.wikipedia.org/wiki/Carlson_curve https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607312177408-FH2441HX6EPDIXJFTXWX/Gibson+Assembly.png The Graduate's Guide to Synthetic Biology https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600213978029-0EFZ74WB38TZMLHRP2GQ/GoldenGate_CorrectAssembly.png The Graduate's Guide to Synthetic Biology Image Credit; https://www.nebiolabs.com.au/applications/cloning-and-synthetic-biology/dna-assembly-and-cloning/golden-gate-assembly https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600215120017-3RQFXC9S64OH879L2XPW/mutagenic+primers.PNG The Graduate's Guide to Synthetic Biology Example: 3 pairs of mutagenic primers for a round-the-world SDM PCR. Note that only 2 primers would be used at a time, these designs were the product of trial and error. Primer set 3 produced the desired product, disabling the start codon of the downstream GFP gene. https://www.neosynbio.com/bufferp2 daily 0.75 2020-12-10 https://www.neosynbio.com/3dprinted-prosthetic-limbs daily 0.75 2020-08-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598503245615-C7AOIM04SDBILX7252EB/60561160_2289355657789366_5721832483444490240_o.jpg 3D-Printed Prosthetic Limbs - First Plastic… Then FLESH 3D-Printed Prosthetic Limbs https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598576994759-R4BXAL3CPO3RVNEW3COS/TelAviv_Heart.jpg 3D-Printed Prosthetic Limbs Image source: https://www.3dnatives.com/en/future-3d-bioprinting1805201741/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598577927793-TZ718P8UU3SRH63P25U5/functionalheart.jpg 3D-Printed Prosthetic Limbs Beating heart grown with stem cells by Massachusetts General Hospital and Harvard Medical school, Image credit: https://www.intelligentliving.co/first-3d-printed-hearts-grown-stem-cells/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598578049479-IH2LBT0PE93I1EED1SDQ/36327649_1805867702804833_562509719273472000_o.jpg 3D-Printed Prosthetic Limbs Model: https://hub.e-nable.org/s/e-nable-devices/wiki/page/view?title=Phoenix+v2+Hand https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598578355322-HHL3PNJN3CUYWBGHDMCL/Inkjet_Bioprinting.jpg 3D-Printed Prosthetic Limbs Image credit: https://www.3dnatives.com/en/future-3d-bioprinting1805201741/ https://www.neosynbio.com/buffern3 daily 0.75 2020-12-10 https://www.neosynbio.com/tips-tricks daily 0.75 2020-11-03 https://www.neosynbio.com/bufferp1 daily 0.75 2020-12-10 https://www.neosynbio.com/bufferpb daily 0.75 2020-12-10 https://www.neosynbio.com/purification-of-plasmid-dna-miniprep-aka-ethanol-lysis daily 0.75 2020-11-05 https://www.neosynbio.com/boil-purification-genomic-dna daily 0.75 2020-11-05 https://www.neosynbio.com/the-sterile-workspace daily 0.75 2020-12-10 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606247462943-PKF2I5GAXMWDB71YJ9BY/Bunsen.png The Sterile Workspace https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607570153014-TTW3BL5V4XJQDZIT9EWN/Laminar+Flow+Hood.png The Sterile Workspace https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607570523117-8QD24Z0HXOUIIOFK6U6R/Biosafety+Cabinet.png The Sterile Workspace https://www.neosynbio.com/purification-of-genomic-dna daily 0.75 2020-08-19 https://www.neosynbio.com/lb-liquid-media daily 0.75 2020-11-25 https://www.neosynbio.com/diy-buffer-mixing daily 0.75 2020-08-17 https://www.neosynbio.com/elution-buffers-te-eb-water daily 0.75 2020-12-10 https://www.neosynbio.com/miniprep-buffer-mixing daily 0.75 2020-07-28 https://www.neosynbio.com/buffer-pe-wash-buffer-2 daily 0.75 2020-12-10 https://www.neosynbio.com/host-selection-criteria daily 0.75 2020-12-08 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1595917262465-3OPP63RETSC2DXTL685G/Hosts+and+Vectors+1.PNG Host Selection Criteria Page 3/5 of the OGTR’s “Types of dealings with GMOs classified as Exempt Dealings” https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1595917423616-J711SP2MESU00LKI4GOC/Hosts+and+Vectors+2.PNG Host Selection Criteria Page 4/5 of the OGTR’s “Types of dealings with GMOs classified as Exempt Dealings” https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597801158924-H4AP69YP4DGQ6TLHXTFJ/glowrabbit.png Host Selection Criteria https://www.neosynbio.com/culturing-conditions daily 0.75 2020-12-09 https://www.neosynbio.com/plasmid-insertion daily 0.75 2020-12-07 https://www.neosynbio.com/sterile-technique daily 0.75 2020-12-10 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606258546349-7RZ0IEG7AB6E2Z22YZK5/Sterile+Technique+for+Isolated+Colonies.png Sterile Technique https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596499314231-PBXNEPYEQVJE0QAPNOIZ/turntables.PNG Sterile Technique https://fac.ksu.edu.sa/sites/default/files/140_mic_10_th_lab.pdf https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597731659951-KSYVM61K2JGVD1SN6A1Q/IMG_20190905_140715.jpg Sterile Technique Left: Concentrated by centrifuging after heat shock protocol. Right: Unconcentrated sample after heat shock protocol. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597731958245-JIOTLDPMP0ZAEXL77V1A/IMG_20190423_082545.jpg Sterile Technique https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597732009791-K4FNWSHTB5MIP0MAHGJ7/IMG_20190418_142811.jpg Sterile Technique https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597732034210-HLVP49R5PMF2DFBBQZNK/IMG_20190418_142838.jpg Sterile Technique https://www.neosynbio.com/antibiotic-stock-solution-mixing daily 0.75 2023-07-12 https://www.neosynbio.com/mixing-media-and-pouring-plates daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606096781276-VTQ939LGFMJE1UVO5E6V/Agar+Plate+Drying.png Mixing Media and Pouring Plates https://www.neosynbio.com/lb-agar-media daily 0.75 2020-11-03 https://www.neosynbio.com/genomic-dna-extraction-with-ctab daily 0.75 2020-12-15 https://www.neosynbio.com/cacl2-chemically-competent-cells daily 0.75 2020-11-05 https://www.neosynbio.com/electroporation-of-electrocompetent-cells daily 0.75 2020-11-05 https://www.neosynbio.com/heat-shock-transformation-of-chemically-competent-cells daily 0.75 2024-05-07 https://www.neosynbio.com/rbcl-chemically-competent-cells daily 0.75 2020-11-09 https://www.neosynbio.com/electrocompetent-cell-preparation daily 0.75 2020-11-05 https://www.neosynbio.com/agarose-electrophoresis daily 0.75 2020-11-30 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606696382879-JY4HSO85MXZ8RBNM815I/Casting+Agarose+Gels.png Agarose Electrophoresis https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597116674595-5S7MS503II7WH5675U5R/32207602_1750046595053611_7302529058535899136_o.jpg Agarose Electrophoresis A 35 cycle PCR of two insulin genes that I was excited to try purify… 100bp Ladder (Lane 1) is extremely faint in ambient light. GelGreen on an IORODEO blue light transilluminator. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597117476740-I3MWQGMUJCRK3VFO8LV2/forwebsite.PNG Agarose Electrophoresis Image Credit: Sebastian S. Cocobia as he works on his Infinite Discovery Machine: https://docs.google.com/document/d/10Y4NgXjMRvG_Vd5D4o2lPJp1ehY2R516gpfrh2PZWfw/edit https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597118098158-ZCAP61UCMYXS3BCVBMT6/notch.PNG Agarose Electrophoresis Test strip of Agarose Gel https://www.neosynbio.com/restriction-enzyme-digest-of-plasmid-and-insert-dna daily 0.75 2020-11-05 https://www.neosynbio.com/ligation-troubleshooting daily 0.75 2020-11-05 https://www.neosynbio.com/flanking-restriction-site-addition daily 0.75 2020-11-09 https://www.neosynbio.com/gradient-pcr daily 0.75 2020-12-07 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596685241974-UTINQAM2S8XJMWXETROC/86766319_2839231162801810_6347751562277814272_o.jpg Gradient PCR Gel was run at 200V and thus has a ‘smile’. All products are actually of identical size, the curve is a relic of impatient electrophoresis. https://www.neosynbio.com/polymerase-chain-reaction daily 0.75 2022-06-15 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596604510842-PXOOYECJ1P5XRSL5I186/69744899_2489911261067137_7835579326543167488_o.jpg Polymerase Chain Reaction The Polymerase Chain Reaction (PCR) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606344932078-B4ZVTF3W0M9PRTTZQ5WZ/Polymerase+Chain+Reaction.png Polymerase Chain Reaction https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606359531242-E5B3Q1EFPE3C87L4FOQJ/Polymerase+Chain+Reaction+Cycle.png Polymerase Chain Reaction Repeated cycles increase the ratio of perfect:imperfect copies of the target region https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596602828207-OW1PBF5ALRI18WI6J8LG/61082075_2308962962495302_8589699966526554112_n.png Polymerase Chain Reaction https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596602869366-VS6BMI13FJ69B9XRSY3C/86766319_2839231162801810_6347751562277814272_o.jpg Polymerase Chain Reaction https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596602939793-YU1T0GZYRQ0B5E5BO4SI/thermocyclertest.png Polymerase Chain Reaction https://www.neosynbio.com/gel-illumination-photography daily 0.75 2020-12-07 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596755713993-KKHN0I6YSJKEZWHWX4MV/etbr+spectrum.PNG Gel Illumination & Photography https://www.thermofisher.com/order/catalog/product/15585011#/15585011 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596759630207-ZALDCYG1BTMPQ2U2V0KM/thiazole+orange.jpg Gel Illumination & Photography https://onlinelibrary.wiley.com/doi/pdf/10.1002/cyto.a.22065 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596759534926-7NZFB07L4G01QKGNZHOI/800px-SYBR_Green_I_spectra.png Gel Illumination & Photography https://commons.wikimedia.org/wiki/File:SYBR_Green_I_spectra.png https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596760491761-JQIKLNCHS7I4CKGHTNOK/32207602_1750046595053611_7302529058535899136_o.jpg Gel Illumination & Photography https://iorodeo.com/products/mini-led-transilluminator This was the first transilluminator that I ever bought and it can give surprisingly clear results (even in ambient light) without breaking the bank. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596760593007-DYCZ639QVUUVZ92HXT7J/87480863_2858195094238750_5711241875776602112_o.jpg Gel Illumination & Photography https://docs.google.com/document/d/10Y4NgXjMRvG_Vd5D4o2lPJp1ehY2R516gpfrh2PZWfw/edit I built according to the specifications in the Open Lab Notebook by Sebastian - but with the wrong wavelength of bulbs, wasting a huge amount of time! Don’t be me! https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596761045038-S1YOFW6BEQ6XVBGW2OFF/H1391.png Gel Illumination & Photography https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596761437559-B8CEF3QKO8806U8K9QXB/86932316_2837287829662810_8622763075742728192_o.jpg Gel Illumination & Photography TFW someone buys you polarised glasses and your eyes are finally safe. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596761326771-BT4RLFP8CELYQPXCO0RA/IMG_20191101_094852.jpg Gel Illumination & Photography My DIY UV imaging enclosure ft. smartphone camera. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596761948489-CU0GYCLKH1ED7OE4ZRQO/38875542_1876758199049116_6055737066535256064_n.png Gel Illumination & Photography This is an old gel image from when I used to use a Chemidoc XRS+ to image my gels. This is probably the best image I ever got from that old machine. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596762064630-6ZQ8I3D6EE7AQ7GB8FGX/sebgel.PNG Gel Illumination & Photography I’ve left in the bulbs for effect, Sebastian normally crops them out. Nonetheless, you can clearly see absolutely everything you need to on this gorgeous gel. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1596759276273-CXBGYPV45KKKSWLMIT3W/Spectrum_multiple_color_filters.jpg Gel Illumination & Photography https://www.vision-doctor.com/images/stories/filter/farbfilter/Spectrum_multiple_color_filters.jpg https://www.neosynbio.com/primer-design daily 0.75 2020-08-19 https://www.neosynbio.com/confirmation-of-edits daily 0.75 2020-11-26 https://www.neosynbio.com/external-sequencing-prep daily 0.75 2020-11-24 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606257752214-JYH2B30IJ3B5S50QBGQY/Flanking+Primers.png External Sequencing Prep https://www.neosynbio.com/golden-gate-assembly daily 0.75 2020-11-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598327262888-YH5X7VBOXJFWZ4A0FFZ6/GoldenGate_CorrectAssembly.png Golden Gate Assembly Image Credit; https://www.nebiolabs.com.au/applications/cloning-and-synthetic-biology/dna-assembly-and-cloning/golden-gate-assembly https://www.neosynbio.com/patch-plate-colony-pcr daily 0.75 2020-11-26 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606255223423-UPNJOOX7KDRPLJ2VT7I1/Flanking+Primers.png Patch Plate & Colony PCR https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606256903863-36TQ4JXCR77V1USRA98X/Patch+Plate.png Patch Plate & Colony PCR https://www.neosynbio.com/restriction-digest-analysis daily 0.75 2020-08-21 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597029287832-RZVDSJPUNDGK89SBOKFK/simulate+gel.PNG Restriction Digest Analysis Here is an example of how you might simulate your gel before actually running it. It’s good practise to pre-register your hypothesis, no matter what area of science! https://slideplayer.com/slide/4567991/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597028778372-5LRVT9NPN3XYG3TY6SX0/T--Sydney_Australia--Results3.png Restriction Digest Analysis Figure 3: Double digest of constructs in pSB1C3 using EcoRI and PstI. Products were run on a 1.5% agarose gel in 0.5X TBE at 80V for 2 hours and post stained with GelGreen (Bio-rad) according to manufacturers protocol. http://2017.igem.org/Team:Sydney_Australia/Results https://www.neosynbio.com/gibson-assembly daily 0.75 2021-08-17 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606439761054-TYY9KRNT5WROF7BWVZ6V/Gibson+Assembly.png Gibson Assembly This is a one-tube-reaction if you ignore the two reactions required to produce the fragment and plasmid with the correct flanking sites (A + B). https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597358404393-5B2LEN50JJC2J6QWF1LJ/example+gibson.PNG Gibson Assembly The plan is to insert the gene on the right (Proinsulin G. Block) into the plasmid on the left (pUS24X), in between MCS1 and MCS2 (multi cloning sites) replacing the amilCP gene. Don’t squint too much, we’ll zoom in on the important annotations. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597367916567-8ZNZPWUT4JNS6JM938DQ/Gibson+Assembly+Map.png Gibson Assembly We can ignore the rest of the plasmid backbone, focussing on the junctions flanking our gene. Lets zoom in on the left junction. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597368336225-62VP1YM2GCD4CNM9EDMV/hotpink.PNG Gibson Assembly In the hot pink, you can see where the two DNA parts will join together at the left junction after the Gibson reaction. It will look similar for the right junction, albeit the sequences encoded will be different. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597369240998-03P4DCIBPMLPK90A0VB3/3%27clamp.PNG Gibson Assembly This is the part that will bind to your gene when we try to amplify it. We will now add a 5’ sequence homologous to our backbone to ensure scarless cloning. Note the Tm at this stage, as it will be more relevant than the Tm of your finished primer. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597369988037-31AM33HCVIZ8C66DJ8OD/Gibfwd1.PNG Gibson Assembly This is the first forward primer (Fwd 1). When used to amplify the gene during PCR, it will add a compatible 5’ end for gibson assembly with your intended plasmid. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597370818959-ESGA2M6ZEZ89GX5IGCFJ/bothprimers.PNG Gibson Assembly The Reverse Primer 2 (Rvs 2) will be used in a separate PCR reaction (with Fwd 2), to amplify the plasmid so that it has compatible ends with your gene. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597374089498-8B43169R4EO32LLENTS1/reverse+complement.PNG Gibson Assembly Same design rules apply. Keep in mind that the #1 primers go together and the #2 primers go together https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597374833738-1VQB3QMV21Y7OVGBHKYL/gibsonbiobrick.PNG Gibson Assembly The Biobrick Prefix and Suffix, originally intended for RE digest, are now replaced with homologous regions to pUS24X. This is a surprisingly simple adaptation of an older Gblock to a newer methodology. Imagine if it was this easy for computer coders… https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597374989051-K3K48A8WAF7EKMLQNXNC/Backbone.PNG Gibson Assembly We’ll amplify all the useful parts of the backbone, while adding on our homologous ends and cutting out the amilCP gene. This will be a significantly longer thermal cycle than the gene, but you may be performing Gibson Assembly with relatively equal size parts. https://www.neosynbio.com/cotton-wool-gel-purification daily 0.75 2020-11-26 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597116401882-2NK6SXUWJWZBVLYZHG0D/cotton+wool.jpg Cotton Wool Gel Purification https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3283835/figure/F1/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597202002345-UCNRWCX07MVMGVHBFRQT/tileshop.jpg Cotton Wool Gel Purification https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3283835/ https://www.neosynbio.com/buffer-qg-gel-purification-buffer daily 0.75 2020-12-10 https://www.neosynbio.com/heat-inactivation-of-restriction-enzymes daily 0.75 2020-08-21 https://www.neosynbio.com/buffer-qg-gel-purification daily 0.75 2020-08-21 https://www.neosynbio.com/column-purification-of-restriction-enzyme-digest daily 0.75 2020-08-21 https://www.neosynbio.com/periplasmic-lysis-with-chloroform daily 0.75 2020-11-26 https://www.neosynbio.com/centrifuge-filter-histag-purification daily 0.75 2020-11-18 https://www.neosynbio.com/induction-of-protein-expression daily 0.75 2020-12-08 https://www.neosynbio.com/cell-lysis daily 0.75 2020-12-08 https://www.neosynbio.com/sds-page daily 0.75 2020-11-18 https://www.neosynbio.com/beadbeating-cell-lysis daily 0.75 2020-11-18 https://www.neosynbio.com/car9-purification daily 0.75 2020-08-17 https://www.neosynbio.com/histag-purification daily 0.75 2020-08-20 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597631820841-LBK7RUXXDP8WIRZ6PCYD/His-IMAC-binding_2148-mod_T.gif His-Tag Purification https://www.takarabio.com/about/bioview-blog/tips-and-troubleshooting/when-your-his-tagged-constructs-dont-bind https://www.neosynbio.com/dna101 daily 0.75 2020-12-11 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603765030502-9DW182J7U6OOIDNAZGPQ/Dna3.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602631729950-SEPSEJQFUU86TGRAOJY9/Transcription.jpg DNA101 The Central Dogma of Molecular Biology. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607640897259-B15QDYA2HJCPI6GHDFJV/Photo_51_x-ray_diffraction_image.jpg DNA101 The legendary “Photo 51” that led to the discovery of the Double Helix. Source: http://scripts.iucr.org/cgi-bin/paper?S0365110X53001939 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602801072459-4PXAAK8J1SHBUFCTT8L3/Meiosismitosis.jpg DNA101 Depending on their location in the organism, Eukaryotic cells (such as your own) will perform either Mitosis or Meiosis. Mitosis will produce more somatic (body) cells and meiosis will produce gamete (sex) cells. This diagram helps illustrate the similarities between the two processes - but keep in mind that Meiosis II has more steps than this diagram is letting on. Keep reading for more details on each… https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602810977170-X3XFT6WREMWYDTICHU9C/binary.jpg DNA101 Plasmids are extra-genomic code that carry their own origin of replication (pictured in red). The origin of replication is where the replication of DNA begins. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597963818596-NYZXOG0OCJYRKMMQGYHD/7.jpg DNA101 Hey look it’s you! Or someone that looks a lot like you… Image credit: https://genomicmedicineuk.com/the-human-genome-structure-and-organization/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603061411462-LVKJXPDHTFWHLRPQDYPF/histone+core.jpg DNA101 How do you fit 46 ~5cm long strands inside a nucleus with a diameter of 5 µm? Wrap it reallllllyyyy tightly. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603074198590-W52A1E937XLM8KTP3GAF/Cell%2Blines.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1597972504181-4ZSKZJR4ZZ8MFXFDE7Q5/proteome.jpg DNA101 Once the Human Genome Project was done, it was time to map the Human Proteome (all proteins and their interactions). Then maybe there might be some clarity as to how humans function… right? RIGHT? Image Credit: https://guardianlv.com/2014/05/proteome-map-nearly-complete/ https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603846021334-93BYNS59P43SQ5AGVJPJ/degrees.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600636988227-TOQBRDEY68OND28Z5FBK/protospacer.png DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603746675164-6OA2OWKNTL1I55JX8CA0/purines+pyrimidines.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603755635735-E6ZKGJ86JQS81J1X9X42/purines+pyrimidines+numbered.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603748597426-QX04POHBZW90AAEE9K9I/Phosphodiester+Bond.jpg DNA101 The complementary strand for this section would have the code TT. Also note that the dotted lines will have likely have phosphates attached, continuing the chain in both directions. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603846935595-28QSS7AWGBAHI3MUBBN0/Hydrogen+Bonds.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603847249763-DM441UK0FERR9QDPT7H2/DNA+Full+Correct.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603847580455-MO0FLPO2KHH23WUAM69C/9bp.PNG DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603849251028-D2ZD1KE60227KUP19L1Y/3d+helix.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603849398484-UVLTHUT5CFWNSE3D4RY1/1+dgree.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603848654846-QYQHKLIY33RBDGTUM3QT/Complex.jpg DNA101 Wrap that DNA in a nucleus and surround it with proteins and cytoplasm and it is beyond our capacity to model in full. It’s also beyond my capacity to draw. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600650730005-M6P14SRXQGLGY0SQ8SWD/6waystoread.jpg DNA101 While there are 6 possible reading frames, only the highlighted orange section is an Open Reading Frame. There is a stop codon, but it’s much later in the sequence. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603772992451-NHPBMLG8JCZ91ZTJE3HZ/trna+synthetase+diagram.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600651442789-NLZKC6RGVM5DKE8HRMWS/mrna-codon-wheel1237881-prints.jpg DNA101 Image source: https://society6.com/product/mrna-codon-wheel1237881_print https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603937026046-ME7KJBBPKSOS7JH9FZLB/Translation+location+tall.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603939840294-5WJXO8Y5VCLZ1SKUFSKI/Transcription+by+mRNA+pol.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604274228922-MOAVAACTM1DQ8E4LMP30/Intron+removal.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604015566372-V5CDNM6HTUDIL3VJCOW7/Ribosomes.jpg DNA101 S refers to the Svedberg value, which relates to sedimentation in a centrifuge. It tells you a little about their size and almost nothing extra. The roles and functions of the rRNA is complex enough to be beyond the scope of this guide. Check out the RNA101 guide and there will hopefully be more info there! For now it can be summarised; rRNA helps the Ribosomal proteins assemble in the correct order, then bind and move the messenger RNA. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604271045996-RQIQ3S5QZ0BGDY6OC7JE/Ribosome+sites+solo.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606360030004-GEA8RPHK163WNACFODPY/Ribosome+sites+full.png DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604284787937-MK76DN9EFLLSNGXUC0GM/Meiosis+Full.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604876308967-1PBNNN0QPZZ61C8U53YO/Cell+cycle.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604358794730-AOSNQX5KQ12IKSAQJP3M/Mitosis.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604530956232-I63G0LG0BPCV921H76G1/polmerase.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604530504998-WOI230SI07LFIWV1A1SL/binary+fission.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605654155513-HPYELYNIUX7DWKUXD1K9/seesaws.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605834358707-IUP0FTF4NL9JX0FKFFCX/Lac+operon.jpg DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602117371646-B9FL97DVO7HTQCHM7DR0/e0376f58ef7d9cff0024ade7f0935d179dbad789.png DNA101 Image source: https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/regulation-of-gene-expression-and-cell-specialization/a/the-lac-operon https://www.neosynbio.com/affinity-tag-purification daily 0.75 2020-08-20 https://www.neosynbio.com/site-directed-mutagenesis daily 0.75 2020-11-18 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598848519955-N3P7N3ZM2TAJUCAIKR9V/sdm.PNG Site Directed Mutagenesis Only a single base in the start codon will be changed, turning off the expression of the downstream GFP gene. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598848601377-WXJATICFXWXQUGSW6VFN/sfGFP-pSB1C3+Map.png Site Directed Mutagenesis It feels like overkill, but back in the days before CRISPR/CAS9 you had to make an entire plasmid just to change a single base. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598848952043-JT5W69P22B1I0KFUNRB0/mutagenic+primers.PNG Site Directed Mutagenesis Oddly enough, moving the mutated base into the centre of the primer ended up working the best. This is more likely a result of a lack of primer dimers/hairpins, 3’ homology is still more important than 5’ homology. Keep in mind I would only ever use two primers at a time for this reaction, it’s not like LAMP! https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1598918058385-U5LC2OIFQOFZXF26HQAY/Schematic-illustration-of-the-megaprimer-method-for-site-directed-mutagenesis-The.png Site Directed Mutagenesis Image Source: https://www.researchgate.net/figure/Schematic-illustration-of-the-megaprimer-method-for-site-directed-mutagenesis-The_fig1_10624497 https://www.neosynbio.com/gblock-design daily 0.75 2020-10-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599017967383-Y2ZPY7MLFKNU3TLGR9OU/Cpro.PNG GBlock Design Example: A 408 bp GBlock that codes for the production of Human Proinsulin in E. Coli. Post-synthesis enzymatic cleavage of the Arginine (Arg) residues with trypsin will remove the C-peptide, leaving just the A+B chains - Human insulin. But that doesn’t matter right now! Look at all the pretty genetic parts! https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599018543168-JJL4K21OGTREG293SEI8/Gene+of+interest.PNG GBlock Design From Start Codon to the Ochre Stop Codon, this open reading frame should be transcribed and translated if inserted into the correct plasmid/genome. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599019286478-9S50PBF94EIAZEUDMTGW/His+tag.PNG GBlock Design The Arginine is not necessary for most experiments, and may even have been a mistake here. We later replaced the Arg with a TEV protease sequence. The GGS4 Linker is a repeating sequence that will ensure the His-Tag sticks out of the molecule and can be caught on the purification matrix. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599019586625-KE9R8E6ZO16232A7ZKCA/Ecotin.PNG GBlock Design The addition of Ecotin and the Ecotin signal peptide commands the cell to send the protein to the periplasmic space. This is ideal since Insulin requires an oxidative environment to fold correctly. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599096126056-X8CML7G14G3KFF4G1VXY/6waystoread.PNG GBlock Design ORF highlighted in Orange. The code can only be read in one frame, in one direction - thanks to the RBS and start codon. DNA is ALWAYS read 5’ -> 3’, but can be read on either strand and therefore in both directions. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599019065222-UI4QNQ3J9DDMAKYQMCXQ/RBS.PNG GBlock Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599020472021-B6GWDEJFKF3AJEB7LF8Z/Prefix.PNG GBlock Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599020490904-ZX4Y86K36MRM94KQTQO0/Suffix.PNG GBlock Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599017018332-AEKEQXWJXCMWS4F262RI/550px-BioBrick-Prefix-Suffix-pSB1C3-Example.png GBlock Design Image source: https://parts.igem.org/Help:Assembly/Scars - Ahh, the ol’ classic iGEM plasmid. Takes me back… https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599095308631-L657U7ARUQKK53EFKNUM/5%27firstphos.PNG GBlock Design The -(P) represents an extra phosphate on the end of the DNA chain. Keep in mind that there are actually phosphates between all of these molecules, in the ribose-phosphate backbone. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599096383818-33C8ZR4KJ33PF325XUQP/Designbuildtestlearn.png GBlock Design https://www.neosynbio.com/backbone-selection daily 0.75 2020-11-24 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599108470674-S6WMMVSWPEL0S7MSC6OJ/pET-15b+Map.png Backbone Selection Example 1: pET-15b (Click image for file download - view on Snapgene/Benchling/etc) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599187407090-278SU2KV8MATXJUD1J9E/pUS24X+Map.png Backbone Selection Example 2: pUS24X (Click image for file download - view on Snapgene/Benchling/etc) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599435365228-DU4M1OUZ3LZRZCQEXAL2/ori1.jpg Backbone Selection Example 1: ori; high-copy-number ColE1/pMB1/pBR322/pUC origin of replication https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599435417303-54HAHQ6YRKO4VSB63IAE/ori2.jpg Backbone Selection Example 2: pBBR1 oriV; replication origin of the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica; requires the pBBR1 Rep protein for replication. pBBR1 Rep; replication protein for the broad-host-range plasmid pBBR1 from Bordetella bronchiseptica https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599438521280-EIJX6GT2YVPVC333SSKN/AmpR.PNG Backbone Selection Example 1: AmpR; Resistance to Ampicillin conveyed by β-lactamase gene https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599438530892-EMOW3TF1JRHAZ19IP4KY/image-asset.png Backbone Selection Example 2: NeoR/KanR; Resistance to Neomycin & Kanamycin conveyed by aminoglycoside phosphotransferase gene from Tn5. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599519186719-JOICHVX22SCEEFWI9MCJ/MCS1.PNG Backbone Selection The two MCS in pET-15b are not marked as features, but can be seen between the EcoRI-HindIII (MCS1) sites and between the BamHI-XbaI (MCS2) sites. The advantage of spread out MCS such as this is that you can decide which parts of the backbone to use and which to replace with your Gblock. For example an XhoI-HinIII digest will use the full T7 induction system, while an XbaI-EcoRI digest will allow you to replace the T7 induction system with something constitutive (always on) as well as introducing a new RBS. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599519191404-NQL9HD8UULIFWGLQZW72/MCS2.PNG Backbone Selection MCS1 and MCS2 are clearly placed so that any Gblock insertion will replace the blue (amilCP) protein in the correct orientation. This provides a useful visual screen of potentially successful assemblies. This system is less flexible and requires the use of the upstream cumate operon after assembly. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599524754878-XNZN6XHZNW16HN668D5X/Promoter1.PNG Backbone Selection pET-15b promoters: The AmpR and lacI promoters have no surrounding regulatory genes and are always on. The T7 promoter is followed by the lac operator and may not work based upon the folding of the DNA around that sequence. If there is no Lactose/IPTG present, it will not be able to function. This promoter is rather specific to the T7 system and may not function separated from its regulatory framework. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599524770896-QZOB8Y2WX7FLIJXY45PA/Promoter2.PNG Backbone Selection pUS24X promoters: The strong PC promoter occurs twice in pUS24X, however the CymR promoter is constitutive and unaffected by any regulatory genes. The other promoter is between the Cumate Operon Binding Sites and will only function if these sites are not chelated (bonded) by cumate. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599525291805-P61S2GPFR9OBI36X2LCS/Terminator1.PNG Backbone Selection pET1-15b Terminators: The T7 Terminator is rather specific like its promoter. While it may work perfectly should you attempt to repurpose pET15b, it should probably be kept with the rest of the T7 system. There are some useful restriction sites on either side of the terminator that allows for some flexibility. While every transcribed gene on the plasmid should have a promoter and terminator, not everyone remembers to annotate properly. As such there are a couple of unannotated terminators on the pET-15b plasmid, perhaps you could try track them down using BLAST! https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599525410594-0CWM28BAZDK1OH37XCOY/Terminator2.PNG Backbone Selection pUS24X Terminator This terminator is rather cleverly designed to work for the existing constitutive expression of the blue protein and to act as a terminator for your gene of interest after insertion. AND it is acting as the terminator for the antibiotic resistance gene that is running in the opposite direction. Brilliant! https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606257628214-A04UNDZTCW3U4MU1CKFL/Lac+operon.png Backbone Selection https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599523392228-8FP8R66SLP4VBY3FEDWI/operon1.PNG Backbone Selection lacI (lac repressor) is always on, and binds to the lac operator. Lactose/IPTG compete for binding and free up the promoter for the RNA polymerase to bind. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1599523417092-MCISQH4S627PO71P4FN9/Operon2.PNG Backbone Selection The CymR (cumate repressor) is always on and binds to the CuO (Cumate Operon Binding Sequence) sites, potentially forming a hairpin structure, but certainly blocking the promoter from functioning. The addition of Cumate (Cumic Acid) https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606095593982-WCBQUPTRATUDOMNAW3YW/Plasmids+on+an+Agarose+Gel.jpg Backbone Selection https://www.neosynbio.com/crisprcas9 daily 0.75 2020-11-26 https://www.neosynbio.com/bunsen-burner daily 0.75 2020-12-08 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600825944123-FPGPLFSIFAHQJWTJMQXZ/Bunsen.crop_598x448_0%2C122.preview.jpg Bunsen Burner Image credit: https://www.tes.com/teaching-resource/forces-and-energy-lesson-14-chemical-potential-energy-bunsen-burner-lesson-11403512 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600825431160-1ODN0VDX2LW7FHIY8UAM/eis0093btl.jpg Bunsen Burner Image credit: https://www.teaching.com.au/product/EIS0093BTL https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600825549082-7PSNFO8K6DC4DWQ8APCW/kemper.PNG Bunsen Burner Image credit: https://www.southernbiological.com/portable-bunsen-burner/ https://www.neosynbio.com/micropipette daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600399860484-BXCTNUU1DVTFALS1MCMW/69484976_950266078640219_1643356677606473728_o.jpg Micropipette https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600402743625-9VECTBPO2FFZT9CF50KH/34672352_1777969902261280_6741294137095487488_o.jpg Micropipette I got about half-way before I realised it requires a central metal tube and I am still yet to source one… https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606700554918-MTYPRJ2T6OTKDXG0X7D0/Reading+Micropipette+Values.png Micropipette https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600402323310-78KP53WHQ260K3S4X17F/Micropipette.png Micropipette https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600402194980-WUQQE13JTNLQWG7Q1OGK/600px-Gilson_schematic.svg.png Micropipette Image credit: https://en.wikipedia.org/wiki/Air_displacement_pipette https://www.neosynbio.com/gloves daily 0.75 2020-09-18 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600388093435-UWI57J9JCNYQGW2H46U1/79924367_2702259549832306_7117383652744364032_o.jpg Gloves https://www.neosynbio.com/inoculation-wand daily 0.75 2020-09-18 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600398872482-7R0LE26QIKNOJI7BIP4S/is-it-safe-to-use-Bunsen-burner-in-a-biological-safety-cabinet.jpg Inoculation Wand https://www.neosynbio.com/lab-coat daily 0.75 2020-09-18 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600388967864-0KCC4GVJ3BN9B5DGY8Y4/5dolla.jpg Lab Coat https://www.neosynbio.com/goggles daily 0.75 2020-09-18 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600398478690-91YHQOWHN3LA87BS9UT3/image-asset.jpeg Goggles https://www.neosynbio.com/tools-template daily 0.75 2020-09-28 https://www.neosynbio.com/rna101 daily 0.75 2020-09-21 https://www.neosynbio.com/proteins101 daily 0.75 2020-09-21 https://www.neosynbio.com/plasmids101 daily 0.75 2020-10-19 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602460302421-OLSPYBHK13YM4LUR4VZ7/1-s2.0-S0147619X18301513-ga1.jpg PLASMIDS101 Image Source: https://ars.els-cdn.com/content/image/1-s2.0-S0147619X18301513-ga1.jpg https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1602719245063-0ZS4JE97ITP2H83R9GRA/image-asset.jpeg PLASMIDS101 Image source; https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4480603/#:~:text=Supercoiling%2C%20Linking%20Number%2C%20and%20Linking,molecules%20were%20isolated%20from%20cells.&text=Form%20I%20is%20also%20called,molecule%20is%20closed%20on%20itself. https://www.neosynbio.com/romilliq-water daily 0.75 2020-09-23 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600827412449-PO8WBQLIPQ93GPXVYQ7N/milli-q_advantage_a10_b.jpg RO/MilliQ Water Image source; https://www.labrepco.com/product/milli-qa-reference-water-purification-system/ I’ve never needed water this fancy, RO is fine! https://www.neosynbio.com/ph-meter daily 0.75 2020-09-23 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600829432006-C0RLM4T66QSPQU7U391H/IC860031.jpg pH Meter Image Source: https://www.instrumentchoice.com.au/instrument-choice/environment-meters/ph-meters-2 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1600829355845-OTHFT9C3IINWSG0YF4JY/CWC-Convergent-water-control-pH-solutiojn-calibration-ph-4-7-10-250-large.jpg pH Meter Sauce: https://cwc.com.au/news-events/how-to-store-and-use-ph-buffer-solutions-for-calibrating-ph/ https://www.neosynbio.com/silica-spin-columns daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601339229462-E1JF28J5TGQJHA6PSBIX/49825983_2093085314083069_6283794027452039168_o.jpg Silica Spin Columns The Miraprep protocol allows for some pretty insane purification yields. https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601339481556-ZUJBG3T3VMNBZFAFIWJ7/41598_2018_23766_Fig1_HTML.jpg Silica Spin Columns Miniprep can occur via vacuum pressure or via centrifugation. https://www.nature.com/articles/s41598-018-23766-9 https://www.neosynbio.com/disinfectant-spray daily 0.75 2020-09-27 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601250608606-HV50AU7CS1I7086R9Z2M/image-asset.jpeg Disinfectant Spray https://www.neosynbio.com/petri-dishes daily 0.75 2020-09-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601250997284-7X04DISMPMBJ4G6YY31G/image-asset.jpeg Petri Dishes https://www.neosynbio.com/tubes-tube-racks daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601252267256-TTDV6ORXRZ5N4NA8WLV7/IMG_20200401_130616.jpg Tubes & Tube Racks https://www.neosynbio.com/fridges-freezers daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601271539337-CZXPE4LWEGRLG644S449/70928786_2530275270364069_7666269843129106432_o.jpg Fridges & Freezers Anyone who caused a reagent to enter drawer 4 was officially in my bad books. https://www.neosynbio.com/elisa daily 0.75 2020-10-01 https://www.neosynbio.com/pipetteboi daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601417142358-2KRBX7WXSUO0UTRF4YT9/Pipeboy-acu-2-Showcase.jpg Pipetteboi https://www.neosynbio.com/weighing-scale daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601414759796-D28AKMYYZXPWECICFWVE/0-02-07-a186682c71fe6ac1af68bba287f915b50f603928f587c4f14aba81feb2359289_eaba7a34.jpg Weighing Scale https://www.neosynbio.com/glassware daily 0.75 2020-12-09 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601416189967-RK83HPK56GGUYSLLB17K/IMG_20191216_120843.jpg Glassware https://www.neosynbio.com/microwave daily 0.75 2020-12-08 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601335548134-A6XNE4RNE01BA6MYZZ0I/image-asset.jpeg Microwave https://www.neosynbio.com/liquid-waste daily 0.75 2020-09-27 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601249971417-IIB38T3E9DNJQC6Z4OHX/vials-1781316_960_720.jpg Liquid Waste https://www.neosynbio.com/incubator daily 0.75 2020-09-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601327830813-SDBQP7CXK5418G03PCNU/MVIMG_20190409_102004.jpg Incubator https://www.neosynbio.com/centrifuge daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601336592697-I665C5QEFY5H8KFPV2X9/centrifuge+5424r+eppendorf.jpg Centrifuge Image credit: https://www.biocompare.com/Product-Reviews/188027-Centrifugation-of-RNA-and-protein-samples-at-4-degree-Celsius/ https://www.neosynbio.com/autoclave daily 0.75 2020-09-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601252922898-NJIQA7K9JUL98R1F2WZI/IMG_20200513_102728.jpg Autoclave https://www.neosynbio.com/gel-tray-bath daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601419053575-W5T01AGN9YF21SU68IOR/33921060_1769725416419062_7881852548276027392_o.jpg Gel Tray & Bath https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601419139622-7034FYH2NH1CBL9WOAE6/32384111_1754015604656710_9176850467227959296_o.jpg Gel Tray & Bath https://www.neosynbio.com/dc-power-supply daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601340451625-ELJ6M42BESURKXO2LE6K/49175166_2081528401905427_7542656853239398400_o.jpg DC Power Supply https://www.neosynbio.com/heat-block daily 0.75 2020-09-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601328680789-FPKMT8N9ZTPRAVKH4EQU/IMG_20200130_115605.jpg Heat Block https://www.neosynbio.com/hot-water-bath daily 0.75 2020-09-28 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601329230521-B1RRJ846ZVCT947H1151/growgrow.PNG Hot Water Bath https://www.neosynbio.com/illuminator daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601419873260-HECYZ3A17OEOZ8JP4WD1/unnamed+%281%29.jpg Illuminator Chemidoc XRS+ Transilluminator from BioRad. Image source: http://www.marywood.edu/science/labs-equipment/equipment/equipment-detail.html?id=5608b433-b756-4b02-8a9f-fc0e672b24ce https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601419789252-FJY7X35OUBMR7INJKLL6/87480863_2858195094238750_5711241875776602112_o.jpg Illuminator DIY Gel Cisilluminator designed by Sebastian S. Cocobia Design notes: Flowers for Everyone https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601420225965-XANJWE3R07KHXUGHPOR0/32207602_1750046595053611_7302529058535899136_o.jpg Illuminator IoRodeo mini LED transilluminator: https://iorodeo.com/products/mini-led-transilluminator https://www.neosynbio.com/thermocycler daily 0.75 2020-09-29 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601339855758-RMC0RNKF7TAH0VUGU39Y/67876307_2441175729274024_148355523387850752_o.jpg Thermocycler https://www.neosynbio.com/imaging-enclosure daily 0.75 2020-09-30 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601426799126-0P6KSMDD8S8D9KEEEPF9/IMG_20191101_094852.jpg Imaging Enclosure DIY 3D printed imaging enclosure that I designed in CAD and then 3D printed in PLA plastic. The aluminium foil is almost definitely overkill, but I couldn’t be certain that the plastic was blocking 100% of the UV radiation. Illuminator is an second-hand Macrovue. https://www.neosynbio.com/sdspage-enclosure daily 0.75 2020-09-30 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601436102111-FL7JCBZ7PQBF7QU31CPQ/SDSPAGE.PNG SDS-PAGE Enclosure Image source: http://www.bio-rad.com/webroot/web/pdf/lsr/literature/Bulletin_6040.pdf https://www.neosynbio.com/bead-beater daily 0.75 2020-09-30 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601430223348-EZ89ZY2B2FNSB7XJYBCO/beads.PNG Bead Beater https://www.neosynbio.com/southern-blot daily 0.75 2020-10-01 https://www.neosynbio.com/antibody-production daily 0.75 2020-10-02 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601598030792-DLLVU4KBWH96TF5O7HFH/image.jpg Antibody Production Image source; https://www.news-medical.net/life-sciences/Types-of-Antibodies.aspx https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601599091946-R75RHFDCATT9E6JS9T23/34605tn.jpg Antibody Production Image Source: https://www.medscape.com/answers/136897-121036/what-is-immunoglobulin-g-igg-deficiency https://www.neosynbio.com/eastern-blot daily 0.75 2020-10-01 https://www.neosynbio.com/cardinal-blots daily 0.75 2020-10-02 https://www.neosynbio.com/knockout-experiments daily 0.75 2020-10-01 https://www.neosynbio.com/genetic-logic-gates daily 0.75 2020-10-11 https://www.neosynbio.com/aptamer-production daily 0.75 2020-10-02 https://www.neosynbio.com/northern-blot daily 0.75 2020-10-11 https://www.neosynbio.com/western-blot daily 0.75 2020-10-11 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1601953522172-V4G5XCXUUIYQRC4XFEOD/GelSandwich.gif Western Blot Image Source: https://openwetware.org/wiki/M465:Western_blots_part_I https://www.neosynbio.com/101-guides daily 0.75 2020-11-27 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603062417931-FNSR7WMJQCSPWZLKDVBP/Dna3.jpg Genetic Theory 101 - DNA101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1603062718373-7NV6TTKLRDLCZTUX0ZGP/pET-15b%2BMap2.jpg Genetic Theory 101 - Plasmids101 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606430405181-WX0IFS258U710QW9HFRS/Flanking%2BPrimers.jpg Genetic Theory 101 - Primer Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606430980223-IOUR69PJ52U1WN8ZJAGI/pUS24X%2BMap.jpg Genetic Theory 101 - BackBone Selection https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606430495492-L22JEES8I1Y6ZL0ENK7D/Lac%2Boperon.jpg Genetic Theory 101 - Gblock Design https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606435319224-DUKV1R1QDP5Z43PYGOAC/Coming%2BSoon.jpg Genetic Theory 101 - RNA101/Proteins101 https://www.neosynbio.com/glossary daily 0.75 2020-11-09 https://www.neosynbio.com/diagrams daily 0.75 2020-11-30 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https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1604891978134-1M940XI4M57LSLO8O24K/trna+synthetase+diagram.jpg Diagrams https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606258671082-XV0FM7XUPBTP3ZQWQEOD/Sterile+Technique+for+Isolated+Colonies.png Diagrams https://www.neosynbio.com/sundry-tools daily 0.75 2020-12-10 https://www.neosynbio.com/laminar-flow-hood daily 0.75 2020-12-10 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606175984288-U2A0K132K7AYVRVRGXR4/laminar+flow+hood.jpg Laminar Flow Hood Image source: https://www.nuaire.com/airegard-nu-140-vertical-laminar-airflow-workstation https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607571537556-LWTQ6C5VU3GNYKTALRES/Laminar+Flow+Hood.png Laminar Flow Hood https://www.neosynbio.com/fplc daily 0.75 2020-11-19 https://www.neosynbio.com/ice-machine-ice-box daily 0.75 2020-11-20 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605833881903-G1HP1LEXXUZ8OTOV2WRC/CX-ims-100-xopen-400x400.jpg Ice Machine & Ice Box Image Source: https://www.hinotek.com/lab/ims-series-snow/ https://www.neosynbio.com/vortex daily 0.75 2020-11-20 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605833106461-8GJP0HRNFVMKYCW970QV/093fbe7ec374f8703be9841130bef486b845b30c.jpeg Vortex Image credit: https://forum.e-liquid-recipes.com/t/lab-grade-vortex-mixer-for-cheap/220013 https://www.neosynbio.com/centrifugal-filter-unit daily 0.75 2020-11-19 https://www.neosynbio.com/consulting-directory daily 0.75 2020-11-25 https://www.neosynbio.com/hplc daily 0.75 2020-11-19 https://www.neosynbio.com/biosafety-cabinet daily 0.75 2020-12-10 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606177585018-JT0ZHDGQ56JMIT4DXPIR/biosafety+cabinet.jpg Biosafety Cabinet Image Source; https://biobasemeihua.en.made-in-china.com/product/WvSEabgUscrm/China-Biobase-100-Air-Exhaust-Class-II-B2-Biosafety-Cabinet-Biological-Safety-Cabinet-Wth-a-HEPA-Filter.html https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1607571693339-28HFQL84XB2IQ0VNMWWU/Biosafety+Cabinet.png Biosafety Cabinet https://www.neosynbio.com/oven daily 0.75 2020-11-22 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606088241869-HN6Z3TW3FL3EWCOVGJWA/image-asset.jpeg Oven Image Source (and great resource): https://megadepot.com/resource/lab-ovens-and-muffle-furnaces-review https://www.neosynbio.com/glass-spreader daily 0.75 2020-11-20 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1605833194635-WQ1MO92SPQTAMF1WURH6/0005876_cell-spreaders-glass.jpg Glass Spreader Image Source: https://www.orioner.com/Cell-Spreader-L-Shaped-Glass-Hockey-Stick-Autoclavable-Non-Sterile https://www.neosynbio.com/shaker daily 0.75 2020-11-19 https://www.neosynbio.com/nanodropqubit daily 0.75 2020-11-22 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606087679036-NRX9KQ2IL1MJ6CMWQZ4T/ND2000008-800.jpg Nanodrop/Qubit Image Source: https://www.labtech.com/thermofisher/nd2000/ https://www.neosynbio.com/spectrophotometer-cuvettes daily 0.75 2020-11-22 https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606085186832-QVG8VXA1LSR4RY06LO14/gb-onda-spectrophotometer1-web.jpg Spectrophotometer & Cuvettes Image source: https://www.chromservis.eu/p/onda-touch-uv-21-spectrophotometer-including-cuvette-holder-and-4-glass-and-2-quartz-cuvettes-usb-flash-drive-and-calibration-report https://images.squarespace-cdn.com/content/v1/5ee69c37561fcb33d73de7cf/1606086270394-5ASVGSUV16STOH9LJEOU/61435162_2315957788462486_7447013540784766976_o.jpg Spectrophotometer & Cuvettes DIY Spectrophotometer, built for me by one of my most talented students. https://www.neosynbio.com/chromatography-columns daily 0.75 2020-11-19 https://www.neosynbio.com/parafilm daily 0.75 2020-12-10 https://www.neosynbio.com/syringe-filters daily 0.75 2020-12-10