Preparing buffers

From Molecular Biology Wiki

How to Prepare Buffers

When preparing buffers:

• Use the highest purity and grade of buffer reagents.
• Prepare solutions with fresh double-distilled water ddH2O or deionized H2O.
• When removing buffer powders / reagents from vials do not use spatulas - carefully shake out amount needed.
• Always wear gloves and a lab coat when preparing buffers.
• Use goggles for eye protection and a mask when required (check the MSDS sheet or read the chemical label for instructions)

Destillation or Deionization units

Using Milipore Water Destillation or Water deionization units for Buffers:

When using millipore (or other purifying units) water, make sure you only use H2O with a resistance of at least 18 MOhm/cm.

Sterilizing Buffers

To autoclave a 0.5 L (liter) solution you should autoclave for 15 minutes at 121°C at 1 bar.

To autoclave a 1 liter solution you should autoclave for 20 minutes at 121°C at 1 bar.

NOTE: You should NOT autoclave buffers containing:

• glucose
• SDS
• β-mercaptoethanol

In this case, filter sterilize solutions through a 0.22 µm filter as an alternative!

See Autoclaving for more information.

pH Measurements

See pH measurement

See preparing RNA buffers and solutions