Table 1. Ligation Efficiency of PCR-Amplified DNA to BamHI Site of pGEX-3X Plasmid Vector.
| Treatment of PCR Products Before Linker Addition |
Number of Clones Analyzed |
Positive Clones |
%of Recombinants |
|---|---|---|---|
| Without kinase reaction a | 8 | ND | ND |
| With kinase reaction b | 6 | 2 | 33 |
Each ligation reaction constituted a 10 μl volume from which 1 μl was used to transform bacterial cells. Competent cells were prepared by CaCl2 method (7).
Reaction a is average of 10 separate experiments.
Reaction b is average of 3 separate experiments.
NO = Not detectable.