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Fig. 3 | BMC Biotechnology

Fig. 3

From: A simple and efficient seamless DNA cloning method using SLiCE from Escherichia coli laboratory strains and its application to SLiP site-directed mutagenesis

Fig. 3

Outline of PCR-based site-directed mutagenesis using SLiCE. Arrows represent PCR primers. a Original overlap extension method for site-directed mutagenesis. b SLiCE-mediated PCR-based site-directed mutagenesis (SLiP site-directed mutagenesis). The linearized vectors were prepared by PCR amplification or by digesting the vector with restriction enzymes. DpnI was active in the PCR buffers. In the case of linearized vectors prepared by restriction enzymes, DpnI added to the insert DNA fragment mixture was heat-inactivated for 15 min at 80 °C

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BMC Biotechnology

ISSN: 1472-6750

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