Buffer N3 (Neutralisation Buffer)

Final Buffer Concentration: 

• 4.20 M Gu-HCl 

• 0.90 M CH3COOK (pH = 4.8)

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!

• CH3COOK - Potassium Acetate - If you’re mixing it yourself, you will need:

  • KOH - Potassium Hydroxide

  • CH3COOH - Glacial Acetic Acid

  • Ice box

Protocol:

For Mixing 200 ml of Buffer:

1. Dissolve 80.25 g Gu-HCl in 100mL Autoclaved dH2O.

2. Add 17.67 g potassium acetate (if mixing yourself: 0.18mol = 10.09 g KOH dissolved in water + 10.809 ml glacial acetic acid).

3. Adjust the pH to 4.8 with glacial acetic acid.

4. Make up to 200 ml with autoclaved dH2O.

5. Don’t need to sterilise.

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

Video: