Features
■ Simple and fast: The kit adopts non-strand substitution plasmid amplification technology. It only needs 4 steps to realize the transformation from wild-type strain to mutant strain, without the time-consuming and labor-consuming steps such as multiple rounds of PCR and sub-cloning. ■ High-efficiency primer: The kit adopts the principle of partially overlapping primer design, so that more mutant plasmids can be obtained by amplification. ■ Widely applicable: The kit can not only carry out single-site mutation, but also multi-site mutation. It can mutate up to 5 sites. ■ Strong adaptability: The kit can carry out site-directed mutation on plasmids with a maximum size of 10 kb, basically covering all commonly used plasmids. ■ High mutation rate: the kit has the function of double digestion of methylated plasmid templates in vitro and in vivo, ensuring higher mutation rate.Site-Mutation Reaction Setup and PCR Program
■ For single primer multi-site mutation, the mutation rate will be lower than that of single site mutation because of the increased number of mutation sites. According to our experimental data, when the number of mutation sites reaches 5, the mutation positive rate will be reduced to 50%. Therefore, in this case, it is recommended to increase the number of clones verified. ■ The kit supports multi-primer multi-site mutation, so that mutation experiments can be simultaneously carried out in a wider range of genes. The upper limit of the number of mutation sites is still 5. ■ It is suggested that the control plasmids and primers supplied in the kit should be applied when carrying out new mutation experiments so as to facilitate analysis of experimental problems.
