FAQ: What is the activity of Salt-T4 DNA Ligase in other buffers?

Salt-T4 DNA Ligase is 100% active in T4 DNA Ligase Buffer supplemented with 100 mM NaCl, NEBuffer 3.1 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 1.12

25%

NEBuffer 1.12 supplemented with 100 mM NaCl

100%

NEBuffer 2.12

50%

NEBuffer 2.12 supplemented with 100 mM NaCl

100%

NEBuffer 3.12

100%

CutSmart® Buffer2

25%

CutSmart 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 CutSmart® Buffer chart.