2-Mercaptoethanol → HCID 131726708, compound

Participants

Reaction Steps

100 μl of a DNA ligase reaction mixture (50 mM Tris-HCl (pH 8.0 at 25° C.); 10 mM 2-mercaptoethanol; 1 mM ATP; 2 μg lambda DNA digested with Hind III; 2 Weiss units of T4 DNA ligase) was added to the prepared reaction tubes. Following incubation of the magnesium oxalate-containing tube with the DNA ligase reaction mixture at 30° C. for t=1, 2, 3, 5 and 10 min., 15 μl samples were removed to clean tubes (without magnesium oxalate) and incubation proceeded for an additional hour at 30° C. Electrophoretic analysis indicated that an incubation time of 10 min at 30° C. was needed to allow enough magnesium to diffuse from the resin in order to activate T4 DNA ligase (FIG. 3, lane 5).

Provenance & Evidence