1M Tris-HCl
Dissolve 121.1g of Tris base in 800 ml of water. Adjust the pH to the desired value by adding concentrated HCl <!> according to the table below.
pH HCl
7.4 ~70 ml
7.6 ~60 ml
8.0 ~42 ml (most common)
Allow the solution to cool to room temperature before making final adjustments to the pH
Adjust the volume of the solution to 1 liter with water
Dispense into aliquots and sterilize by autoclaving
If the 1M solution has a yellow color, discard it and obtain Tris of better quality
20X SSC
Dissolve 525.9g of NaCl and 264.6g of sodium citrate in 2.4L of water
Adjust to pH 7.0 with a few drops of a 12N solution of HCl <!>
Bring the volume to 3 liters with water
Dispense into aliquots and sterilize by autoclaving
The final concentrations of the ingredients are 3M NaCl and 0.3M sodium citrate
Electrophoresis and Gel-loading Buffers
TAE (50X)
Dissolve 242g of Tris base in ~700ml water
Add 57.1ml of glacial acetic acid <!>
Add 100ml of 0.5M EDTA (pH 8.0)
Bring volume up to 1L with water
Dispense into aliquots and sterilize by autoclaving
TBE (5X)
Dissolve 189g of Tris base in ~3L of water
Add 96.25g of boric acid
Add 70ml of 0.5M EDTA (pH 8.0)
Bring volume up to 3.5L with water
Dispense into aliquots and sterilize by autoclaving
TBE (1X)
200ml 5X TBE
800ml water
LB Freezing Buffer
36 mM K 2 HPO 4 (anhydrous)
13.2 mM KH 2 PO 4
1.7 mM sodium citrate
0.4 mM MgSO 4 *7H 2 O
6.8 mM ammonium sulfate
4.4% (v/v) glycerol
100 ml LB broth
Dissolve the salts into 100ml of LB to the specified concentrations
Measure 95.6ml of the resulting solution into a fresh container and then add the 4.4ml of glycerol
Mix the solution well and then sterilize by passing it through a 0.45-ul disposable Nalgene filter
Store the sterile freezing medium at a controlled room temperature (15-25°C)
.5M EDTA (pH 8.0)
Add 186.1g of disodium EDTA*2H 2 O to 800 ml of water
Stir vigorously on a magnetic stirrer
Adjust the pH to 8.0 with NaOH (~20 g of NaOH pellets)
Bring the volume to 1L with water
Dispense into aliquots and sterilize by autoclaving
The disodium salt of EDTA will not go into solution until the pH of the solution is adjusted to ~8.0 by the addition of NaOH
5M NaCl
Dissolve 292g of NaCl in 800 ml of water
Bring the volume to 1 liter with water
Dispense into aliquots and sterilize by autoclaving
Store the NaCl solution at room temperature.
Dissolve 200g of electrophoresis –grade SDS<!> in 900 ml of water
Heat to 68 C, and stir with a magnetic stirrer
If necessary, adjust the pH to 7.2 by adding a few drops of concentrated HCl <!>
Bring the volume to 1 liter with water
Store at room temperature
Sterilization is not necessary, do not autoclave
3M Sodium Acetate (pH 5.2 and pH 7.0)
Dissolve 102.07g of sodium acetate*3H 2 O in 200 ml of water
Adjust the pH to5.2 with glacial acetic acid <!> or adjust the pH to 7.0 with dilute acetic acid
Bring the volume to 250 ml with water
Dispense into aliquots and sterilize by autoclaving