Table 2.
Analysis of the Mutagenesis Efficiency of the Modified PCR-based Site-directed Mutagenesis Method
| Mutated plasmids | Transformed colonies | Silent restriction enzyme digestion | DNA sequencing | ||||
|---|---|---|---|---|---|---|---|
| Plasmids screeneda | Desired plasmids | Mutagenesis efficiencyb | Plasmids screeneda | Desired plasmids | Mutagenesis efficiencyb | ||
| HP-NAPD98A | 1 | 1 | 1 | 1/1 (100%) | 1 | 1 | 1/1 (100%) |
| HP-NAPY99A | 3 | 3 | 3 | 3/3 (100%) | 1 | 1 | 1/1 (100%) |
| HP-NAPY101H | 4 | 4 | 2 | 2/4 (50%) | 1 | 1 | 1/1 (100%) |
| HP-NAPE103A | 40 | 3 | 1 | 1/3 (33%) | 1 | 1 | 1/1 (100%) |
| HP-NAPE103D | 2 | 2 | 2 | 1/2 (50%) | 1 | 1 | 1/1 (100%) |
| HP-NAPE97GY101H | 28 | 4 | 3 | 3/4 (75%) | 1 | 1 | 1/1 (100%) |
aThe plasmids screened by silent restriction enzyme digestion were isolated from randomly selected clones, whereas the plasmids screened by DNA sequencing were randomly selected from the desired plasmids identified by silent restriction enzyme digestion
bMutagenesis efficiency was calculated as the number of desired plasmids out of a number of plasmids selected for screening