Table 2 Comparison of the performance of helper plasmids in enabling introduction of a sequence encoding a C-terminal 3XFLAG tag on RpoD in E. coli K-12 MG1655
Helper plasmid | pKDTS2 | pSLTS |
|---|---|---|
Fraction of colonies that lost the selection marker after double-strand cleavage of the mutation cassette by I-SceI | 14/20 | 17/20 |
Fraction of colonies that had lost the selection marker for which a fragment of the correct size was amplified by colony PCR using primers flanking the gene of interest | 4/8 | 8/8 |
Fraction of colonies for which the correct PCR product was obtained that contained the desired mutation | 4/4 | 8/8 |