A1: The pack contains 4 vials containing a total of 6 ml of 10X E. coli DNA Ligase Reaction Buffer.
Q2: What is the composition of E.coli DNA Ligase Reaction Buffer?
A2: 1X E. coli DNA Ligase Reaction Buffer: 30 mM Tris-HCl, 4 mM MgCl2, 1 mM DTT, 26 µM NAD+, 50 µg/ml BSA, pH 8.0 @ 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.
