Table 2 Ligation reaction for TA cloning.
From: Differential effects of KRAS G-domain and hypervariable region mutants on cancer phenotypes
Dephosphorylation reaction | |
|---|---|
Component | Amount |
empty pTarget vector | 50 ng |
alkaline phosphatase | 1 U |
10X phosphatase buffer | 1 µl |
water | to 10 µl |
Ligation reaction | |
Component | Amount |
Insert DNA | 1:5 molar ratio |
10X ligation buffer | 2 µl |
T4 DNA ligase | 1 U |
10 mM ATP | 1 µl |
water | to 20 µl (total) |
