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Figure 3 | BMC Biotechnology

Figure 3

From: Patch cloning method for multiple site-directed and saturation mutagenesis

Figure 3

Multiple site-directed mutagenesis of a M-MLV RT gene. Schematic presentation of six- (a) or nine-point (b) simultaneous mutation in M-MLV RT gene is shown. DNA fragments amplified by PCR were assembled with the NdeI–XhoI-digested pET16b vector using MUPAC. Length in base pair is shown over each DNA fragment. Plasmids were extracted from each six- and nine-point site-directed mutagenesis experiment. After digestion with NdeI and XhoI, the digested plasmids were analyzed using 1.2% agarose gel electrophoresis (lanes 2–17, the six-site experiments; lanes 19–34, the nine-site experiment). Lane 1 and 18 contain 1000 bp DNA ladder marker. CFU in each experiment are listed in Additional file 1: Table S3. (c) Summary of 6- or 9-points mutation. Mutants were analyzed using electrophoresis and DNA sequencing.

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