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

Table 2 Effect of end homology length on SLiCE cloning

Homology length (bp)	Number of colonies^a		Cloning efficiency		Cloning accuracy (%)^b
Homology length (bp)	Prx IIE	G6PDH1	Prx IIE	G6PDH1	Prx IIE	G6PDH1
10	558 ± 74.5	585 ± 121	11/16	16/16	100.0	93.8
15	875 ± 43.9	777 ± 186	16/16	15/16	100.0	100.0
19	972 ± 162	1,070 ± 289	15/16	16/16	100.0	100.0
24	926 ± 28.6	519 ± 74.8	16/16	16/16	100.0	100.0
29	483 ± 34.8	520 ± 51.4	16/16	16/16	100.0	100.0
34	105 ± 37.2	150 ± 47.1	14/16	16/16	100.0	87.5

^aEach value of “number of colonies” is the mean ± standard deviation of three independent experiments. ^bCloning accuracies are given as the percentage of correctly cloned expression vectors in colony-PCR positive clones. The insert DNA fragments and linearized vector DNA were prepared by PCR. The SLiCE (JM109) reaction was performed for 10 min at 37 °C with 1:1 and 3:1 molar ratios of insert to vector for Prx IIE and G6PDH1, respectively

ISSN: 1472-6750