High efficiency cells to create uracil-containing DNA for site-directed mutagenesis
1 Kunkel T.A. 1985. Rapid and efficient site-specific mutagenesis without phenotypic selection. Proc. Natl. Acad. Sci. 52:488-492.
Lucigen is the sole provider of high-efficiency electrocompetent CJ236 cells. This cell line was created by adding the pCJ105 plasmid to BW313 cells2, and is ideally suited to the Kunkel directed mutagenesis method. This method relies on the cells producing plasmid DNA containing some uracil residues. The dut- and ung- mutations prevent the cells from removing uracil from DNA caused by misincorporation or dCTP deamination. Note that these cells contain an F’ which confers resistance to Chloramphenicol at low levels.
As Electrocompetent cells, the transformation efficiency is 1 × 109 cfu/µg.
[F’ Tra+ Pil+ (CamR)] ung-1 relA1 dut-1 thi-1 spoT1 mcrA
2 Joyce, C.M., N. Grindley 1984. Method for determining whether a gene of Escherichia coli is essential: application to the polA gene. J.Bacteriol. 158: 636-643