Editing 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:&nbsp;400 units/ul,&nbsp;&nbsp; 1.1mg/ml,&nbsp;if&nbsp; 2,000&nbsp;U/ul ==> 6.25mg/ml

'''4 X T4 DNA Ligation Buffer''': 250 mM Tris-HCI, 40 mM MgCl<sub>2</sub>, 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&nbsp;mM ATP solution:

55&nbsp;mg ATP-Na2 in 10 ml H2O

'''500 ul 4 X&nbsp;T4 DNA Ligation&nbsp; Buffer:&nbsp;'''125 ul of 1 M Tris-HCl, pH7.6,&nbsp; 20 ul of 1M MgCl<sub>2</sub>, 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&nbsp;T4 DNA Ligation&nbsp;Buffer:&nbsp;'''250 ul of 1 M Tris-HCl, pH7.6,&nbsp; 40 ul of 1M MgCl<sub>2</sub>, 4 ul of 1M DTT, 400 ul of 10 mM ATP, 300 mg of PEG 6000, adjust to&nbsp; 1ml with H2O

'''10 ml 4 X T4 DNA Ligation&nbsp; Buffer:&nbsp;'''2.5 ml of 1 M Tris-HCl, pH7.6,&nbsp; 400 ul of 1M MgCl<sub>2</sub>, 40 ul of 1M DTT,&nbsp; 55 mg ATP, 3 g&nbsp;of PEG 6000, adjust to&nbsp; 10 ml with H2O

== NEB Sticktogether Buffer, 2X ==

1X Buffer Components<br/>132 mM Tris-HCl<br/>20 mM MgCl2<br/>2 mM ATP<br/>2 mM DTT<br/>15% Polyethylene glycol (PEG 6000)<br/>pH 7.6@25°C[[Category:Buffer Recipe]]