Fast T4 Ligase Buffer

T4 DNA Ligase, 2,000,000 U/ml in storage buffer: 10 mM Tris-HCl, 50 mM KCl, 1 mM DTT, 0.1 mM EDTA, 50% glycerol, pH 7.5 @ 25°C.

200,000 Units: 100 ul of 6.25 mg/ml T4 DNA ligase

NEB T4 DNA Ligase: 400 units/ul,   1.1mg/ml, if  2,000 U/ul ==> 6.25mg/ml

4 X T4 DNA Ligation Buffer: 250 mM Tris-HCI, 40 mM MgCl₂, 4 mM DTT, 4 mM ATP, 30% PEG 6000, pH 7.6 @ 25°C

20 ul reaction: 5 ul of buffer per ul of enzyme, 100 ul enzyme=500 ul Buffer.

10 mM ATP solution:

55 mg ATP-Na2 in 10 ml H2O

500 ul 4 X T4 DNA Ligation  Buffer: 125 ul of 1 M Tris-HCl, pH7.6,  20 ul of 1M MgCl₂, 2 ul of 1M DTT, 200 ul of 10 mM ATP, 150 mg of PEG 6000, adjust to 500 ul with H2O

1 ml 4 X T4 DNA Ligation Buffer: 250 ul of 1 M Tris-HCl, pH7.6,  40 ul of 1M MgCl₂, 4 ul of 1M DTT, 400 ul of 10 mM ATP, 300 mg of PEG 6000, adjust to  1ml with H2O

10 ml 4 X T4 DNA Ligation  Buffer: 2.5 ml of 1 M Tris-HCl, pH7.6,  400 ul of 1M MgCl₂, 40 ul of 1M DTT,  55 mg ATP, 3 g of PEG 6000, adjust to  10 ml with H2O

NEB Sticktogether Buffer, 2X

1X Buffer Components
132 mM Tris-HCl
20 mM MgCl2
2 mM ATP
2 mM DTT
15% Polyethylene glycol (PEG 6000)
pH 7.6@25°C