Buffer PB (Wash Buffer 1)

Final Buffer Concentration: 

• 5 M Gu-HCl 

• 30 % Isopropanol 

  • pH 7.5 (apparently optional)

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)

• Gu-HCl - Guanidinium chloride or guanidine hydrochloride - Seriously nasty stuff! Be extremely cautious when handling, do not inhale!

• Isopropanol - CH3CHOHCH3 aka. Isopropyl alcohol, propan-2-ol - Very flammable! There is no need to be sterile, so turn off that bunsen!

Protocol:

For Mixing 200 ml of Buffer:

1. Dissolve 95.53 g Gu-HCl in 100 ml Autoclaved dH2O

2. Add 60 ml isopropanol

3. Optional: Balance pH to 7.5 with dilute HCl

4. Make up to 200 ml with dH2O

5. No need to sterilise, never autoclave flammable materials.

Video: