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Table 2 Analysis of the Mutagenesis Efficiency of the Modified PCR-based Site-directed Mutagenesis Method

From: Enhanced Mutant Screening in One-step PCR-based Multiple Site-directed Plasmid Mutagenesis by Introduction of Silent Restriction Sites for Structural and Functional Study of Proteins

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%)
  1. 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
  2. bMutagenesis efficiency was calculated as the number of desired plasmids out of a number of plasmids selected for screening