Buffer P1 (Resuspension Buffer)

Final Buffer Concentration: 

• 50 mM Tris-Cl (pH 8.0)

• 10 mM EDTA 100 ug/uL

• RNAse A 

Equipment & Consumables:

• PPE: Gloves, Lab Coat and Goggles (at least!)

• Sterile Workspace

• Sterile Micropipettes and Tips

• Pipetteboi and tips

• Weighing Scale (accurate to to 0.01g)

  • Weigh boats (or folded paper)

• Filters & Syringes OR Autoclave

• Optional: Heated/Stirring Platform

• pH Meter (recently calibrated)

• Graduated Beaker

• Measuring Cylinder

• 4°C Refrigerator

• ddH20 (MilliQ or RO)

• Tris Base - aka. 2-Amino-2-(hydroxymethyl)propane-1,3-diol but really, just call it Tris.

• EDTA.2NA.2H2O - EDTA Disodium Salt Dihydrate (other hydrations are fine)

• HCl - Hydrochloric Acid

• RNase A - Heat sensitive, especially once in solution, keep bottle and solution refrigerated.

Protocol:

For mixing 200 ml of Buffer:

1. Dissolve 1.21g Tris base and 0.75g EDTA-2H20 in 150mL Autoclaved dH20

2. Adjust pH to 8.0 with HCl using a pH meter

3. Adjust volume to 200 ml with dH2O

4. Filter Sterilise or Autoclave your buffer.

5. Add 20mg RNase A. Store at 4°C after this step is performed.

See video for 100 ml and 1L recipes, safety instructions and much, much more;

Video: