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

Hi-T4 DNA Ligase is 100% active in T4 DNA Ligase Buffer.

Like T4 DNA Ligase, Hi-T4 DNA Ligase has reduced activity in NEBuffer 3/3.1 and HiFi Taq DNA Ligase Buffer due to their high salt content.

Buffer

% Activity1

T4 DNA Ligase Buffer

100%

NEBuffer 1.12

100%

NEBuffer 2.12

100%

NEBuffer 3.12

50%

CutSmart® Buffer2

100%

Taq DNA Ligase Buffer2

100%

HiFi Taq DNA Ligase Buffer2

50%

 

1Activity relative to activity on a cohesive end substrate in T4 DNA Ligase Buffer (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.