Paul.xu at 17:30, 15 January 2021

2021-01-15T17:30:20Z

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'''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[[Category:Buffer Recipe]]

Fast T4 Ligase Buffer - Revision history

Paul.xu at 17:30, 15 January 2021