Salt-T4 DNA Ligase is 100% active in T4 DNA Ligase Buffer supplemented with 100 mM NaCl, NEBuffer r3.1 and HiFi Taq DNA Ligase Buffer (when supplemented with ATP). Salt-T4 DNA Ligase has reduced activity in other NEB restriction enzyme and ligase buffers, however, activity in these buffers can be restored when supplemented with 100 mM NaCl.
Buffer |
% Activity1 |
---|---|
T4 DNA Ligase Buffer supplemented with 100 mM NaCl |
100% |
T4 DNA Ligase Buffer |
25% |
NEBuffer r1.12 |
25% |
NEBuffer r1.12 supplemented with 100 mM NaCl |
100% |
NEBuffer r2.12 |
50% |
NEBuffer r2.12 supplemented with 100 mM NaCl |
100% |
NEBuffer r3.12 |
100% |
rCutSmart® Buffer2 |
25% |
rCutSmart Buffer2 supplemented with 100 mM NaCl |
100% |
Taq DNA Ligase Buffer2 |
25% |
HiFi Taq DNA Ligase Buffer2 |
100% |
1Activity relative to activity on a cohesive end substrate in T4 DNA Ligase Buffer supplemented with 100 mM NaCl (25°C for 10 minutes).
2 Supplemented with 1 mM ATP.
Please be sure to use riboATP (NEB #P0756) as deoxyriboATP will not work. To see its % functional activity in CutSmart, and that of other DNA Modifying enzymes in the cloning workflow, refer to the Activity of DNA Modifying Enzymes in rCutSmart® Buffer chart.