2-Mercaptoethanol → HCID 131726708, compound
Reaction equation
C(O)C(N)(CO)CO.Cl.SCCO.P([O:26][CH2:27][C@H:28]1O[C@@H](N2C3N=CN=C(N)C=3N=C2)[C@H:30](O)[C@@H:29]1O)(OP(OP(O)(O)=O)(O)=O)(=O)O.[C:45]([O-:50])(=[O:49])[C:46]([O-:48])=[O:47].[Mg+2:51]>>[C:45]([O-:50])(=[O:49])[C:46]([O-:48])=[O:47].[Mg+2:51].[CH3:30][CH2:29][CH2:28][CH:27]=[O:26]Participants
5
Procedure
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).
Workup
- additionwas added to the prepared reaction tubes
- additioncontaining tube
- customwere removed to clean tubes (without magnesium oxalate) and incubation
- waitproceeded for an additional hour at 30° C
- customindicated that an incubation time of 10 min at 30° C.
Sources & evidence
- ORD record
- ord-7b0831a53e544ea1b3959dc3ff730df1
- Source dataset
- uspto-grants-2007_12
- Patent number
- US07306929B2
Reaction notes
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