50mM NaPO4, 500mM NaCl, 20mM imidazole, 10% glycerol, 10mM Mg2Cl2, 3mM bME , pH 7.2 (To the extract protease inhibittor capsules)
to make 100ml of the buffer: 0.69g of NaH2PO4.H2O, 2.92 g of NaCl, 10 ml glycerol (100%), 0.136 imidazole, 0.095 MgCl2, 20ul of beta Mercaptoethanoal (14.3 M)
50mM NaPO4, 500mM NaCl, 20mM imidazole, pH 7.2
To make 200ml of the buffer: 1.38 of NaH2PO4.H2O, 5.84g of NaCl, 0.272g imidazole
titrate with NaOH to adjust pH
20mM Tris, 500mM NaCl, 500mM Imidazole, 3mM bME, pH 8.0 (for UvsY keep the pH at 7.5)
To make 100ml of the buffer: 0.242g of Tris, 2.92 g NaCl, 3.4 g imidazole, 20ul of bME (14.3 M)
To make 50ml of the buffer: 0.121g Tris, 1.46g NaCl, 1.7g imidazole, 10ul bME pH 7.5
Gp32:20mM Tris, 500mM NaCl,20% glycerol, 2mM DTT, pH 8.0
To make 200ml of this buffer: 0.49 g tris, 5.84g NaCl, 40ml glycerol, 0.0617g of DTT
UvsX:20mM Tris, 300mM NaCl, 1mM DTT
To make 200ml of this buffer: 0.49 g tris, 3.54g of NaCl, 0.03g of DTT
Bsu:20mM Tris, 500mM NaCl, 1mM DTT, 10% glycerol pH 8.0
UvsY:20mM Tris, 500mM NaCl, 1mM DTT, 10% glycerol pH 8.0
To make 500ml of this buffer: 1.21g Tris, 29.2g NaCl, 0.15g DTT, 50ml glycerol
adjust 250ml at pH 8.0 and remaining 250ml to pH 8.0
Reaction Mix(2x): 2mM DTT, 5% PEG-35K (PEG 20K is a better choice), 50mM Tris (pH 8.0), 100mM potassium acetate
To make 20ml of 2X Reaction Mix:
4mM DTT: add 12.3 mg of DTT
10% PEG-20K: add 2 g of the powder
100 mM Tris 8.0: add 0.242 g
200 mM Potassium acetate: add 0.393 g
Make up 20 ml with ddH20, adjust pH to 8.0
Make aliquots of 1-2 ml and freeze at -20C
Mg soln(20x): 14mM MgCl2 (or Magnesium Sulfate)
to make 10 ml of 20x Magnesium chloride solution of 280 mM concentration
add 0.266g of magnesium chlordie in 10ml water
3 mM ATP, 50 mM phosphocreatine
to make 5ml of 10x energy mix:
creatine phosphate disodium tetrahydrate: 0.81785g
10x of enzyme mix will have