B0202 FAQs for T4 DNA Ligase Reaction Buffer, DNA Modifying Enzymes and Cloning Buffers, Intl, NEB
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T4 DNA Ligase Reaction Buffer FAQ

Q1: What is supplied with the T4 DNA Ligase Reaction Buffer pack?
Q2: What is the composition of the T4 DNA Ligase Reaction Buffer?
Q3: How should the NEBuffer be used?
Q4: The buffer arrived thawed; is it and the enzyme still active?

Q1: What is supplied with the T4 DNA Ligase Reaction Buffer pack?

A1: The pack contains 4 vials containing a total of 6.0 ml of 10X T4 DNA Ligase Reaction Buffer.


Q2: What is the composition of the T4 DNA Ligase Reaction Buffer?

A2: 1X T4 DNA Ligase Reaction Buffer: 50 mM Tris-HCl, 10 mM MgCl2, 10 mM Dithiothreitol 1 mM ATP, pH 7.5 @ 25°C. Supplied as a 10X concentrated stock.


Q3: How should the NEBuffer be used?

A3: The buffer must be completely thawed before use. Dilute the 10X stock with dH2O to a final concentration of 1X. Add the water first, buffer next, the substrate and finally the enzyme. For example, a 50 µl reaction should contain 5 µl of 10X buffer with the rest of the volume coming from the substrate, enzyme and dH2O.


Q4: The buffer arrived thawed; is it and the enzyme still active?

A4: The buffers will often thaw when shipped on wet ice. This is often due to the high salt concentration of the 10X buffer. If the ice pack is still solid and the contents are cold to the touch the buffer and enzyme are fine.