Molecular Techniques and Methods

Modification Enzyme Buffers

Copy Right © 2001/ Institute of Molecular Development LLC

5 x BAL 31 Buffer

3 M NaCl
60 mM CaCl2
60 mM MgCl2
100 mM Tris-HCl (pH 8.0)
1 mM EDTA (pH 8.0)
2 x KGB Buffer

200 mM Potassium Glutamate
50 mM Tris-acetate (pH 7.5)
20 mM Magnesium acetate
100 ug/ml BSA (Fraction V)
1 mM 2-Mercaptoethanol
10 x Blunt-End Ligation Buffer

0.66 M Tris-HCl (pH 7.6)
50 mM MgCl2
50 mM DTT
10 mM Hexamminecobalt chloride
2 mM ATP
5 mM Spermidine HCl
Divide the stock solution into small aliquots and store them at -20oC.
10 x Bacteriophage T4 DNA Ligase Buffer

200 mM Tris-HCl (pH 7.6)
50 mM MgCl2
50 mM DTT (Dithiothreitol)
Divide the stock solution into small aliquots and store them at -20oC.
Add ATP at the time of setting up the reaction.
The final concentration of ATP in the reaction should be 0.5-1 mM.
10 x Klenow Buffer

0.5 M Tris-HCl (pH 7.6)
0.1 M MgCl2
10 x Exonuclease III Buffer

0.66 M Tris-HCl (pH 8.0)
66 mM MgCl2
10 X Mung-Bean Nuclease Buffer

300 mM Sodium acetate (pH 4.5)
500 mM NaCl
10 mM ZnCl2
50% Glycerol
10 x Ligation Buffer

0.5 M Tris-HCl (pH 7.6)
100 mM MgCl2
100 mM DTT (Dithiothreitol)
10 X Nick-Translation Buffer

0.5 M Tris-HCl (pH 7.5)
0.1 M MgSO4
1 mM DTT (Dithiothreitol)
500 ug/ml BSA (Fraction V)
Divide the stock solution into small aliquots and store them at -20oC.
Reverse Transcriptase Buffer

50 mM Tris-HCl (pH 7.6)
60 mM KCl
10 mM MgCl2
1 mM of each dNTP
1 mM DTT (Dithiothreitol)
1 unit/ul Placental RNase inhibitor
50 ug/ml Actinomycin D
10 X S1-Nuclease Buffer

2 M NaCl
0.5 M Sodium acetate (pH 4.5)
10 mM ZnSO4
5% Glycerol
10 x CIP Dephosphorylation Buffer

10 mM ZnCl2
10 mM MgCl2
100 mM Tris-HCl (pH 8.3)
10 X Bacteriophage T4 DNA Polymerase Buffer

0.33 M Tris-acetate (pH 8.0)
0.66 M Potassium acetate
0.1 M Magnesium acetate
5 mM DTT (Dithiothreitol)
1 mg/ml BSA (Fraction V)
Divide the stock solution into small aliquots and store them at -20oC.
10 X Bacteriophage T4 Polynucleotide Kinase Buffer

0.5 M Tris-HCl (pH 7.6)
0.1 M MgCl2
50 mM DTT (Dithiothreitol)
1 mM Spermidine HCl
1 mM EDTA (pH 8.0)


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