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

Table 1 The cloning efficiencies using SLiCE from different E. coli laboratory strains

^aNumber of colonies is represented as CFU per nanogram of vector. Each value of “number of colonies” is the mean ± standard deviation of three independent experiments. ^bCloning efficiencies for the insert DNA are represented as “number of clones with the confirmed correct insert length by colony-PCR/number of colonies subjected to colony-PCR”. The insert DNA fragments were amplified using 19-bp overlap primers. The linearized vector DNA was prepared by PCR. The SLiCE reaction was performed for 60 min at 37 °C with an insert:vector ratio of 1:1 and 3:1 for Prx IIE and G6PDH1, respectively

ISSN: 1472-6750