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Figure 3 | BMC Biotechnology

Figure 3

From: MALS: an efficient strategy for multiple site-directed mutagenesis employing a combination of DNA amplification, ligation and suppression PCR

Figure 3

Agarose gel electrophoregram demonstrating the types of molecules forming during one individual round of mutagenesis. Lanes 1 and 2, PCR of target DNA with SO1/IR1 and SO2/IF1 oligonucleotides; Lane 3, homomeric (type A) and heteromeric (type C) DNA molecules formed during ligation of SO1/IR1 and SO2/IF1 DNA fragments (also, see Figure 1); Lane 4, suppression PCR with SO1 and SO2 oligonucleotides. The scheme on the right shows a graphical representation of type A and type C molecules that are present in lane 3 of the gel. During suppression PCR, intramolecular hybridization of inverted repeat sequences prevents binding of SO1 and SO2 oligonucleotides to the type A molecules that prevent their effective replication. Type C molecules amplify exponentially (see Additional File 1 for further details). Lane M, DNA size ladder with indicated positions of 1, 2, 3, 4, 6, and 10 kb DNA fragments (GeneRuler 1 kb DNA Ladder, Fermentas).

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BMC Biotechnology

ISSN: 1472-6750

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