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|Fast: 5 minutes at room temperature.||Simple: only add PCR products.||High efficiency: up to 90% clones with correct insert.|
Partially overlapping primers were used into PCR amplification, and the amplified products were visible by gel electrophoresis. The amplified products were ring-shaped and easy to be transformed.The mutant clones were screened by DMT enzyme digestion in vitro by degrading non-mutant plasmid template (methylated plasmid template) and degrading non-mutant plasmid template (methylated plasmid template) by DMT competent cells in vivo.