SORS: A Universal One-Round PCR-Based Method for Site-Directed Mutagenesis.

@article{citeulike:2770224, abstract = {We have developed a novel protocol for site-directed mutagenesis of double-stranded DNA. The procedure, termed SORS (named because it undergoes the sequential procedure of segmentation-overhang creating PCR-reannealing-splicing) mutagenesis, is exemplified by a substitution, a deletion, and an insertion of nucleotide(s) in target genes. The template DNA is PCR-amplified into two separate segments divided at the prospective mutation site, and each segment is amplified in two parallel PCRs using primers introducing the mutation. The primers are designed to be able to create protruding bases upon pooling, denaturing, and reannealing the two parallel reactions. The protruding bases at the prospective junction of the two segments are mutually complementary; therefore, the two segments can be re-spliced together to generate the mutated gene. Compared to previously published protocols, this procedure is rapid, restriction-independent and ensures higher success rate and lower potential to produce second-site mutations.}, address = {Key Research Lab of Hormone and Development Affiliated to the Ministry of Health, Tianjin Medical University, Tianjin, 300070, People’s Republic of China, blwang7212@yahoo.com.cn.}, author = {Wang, Bao-Li L. and Quan, Jin-Xing X. and Liang, Hui and Jiao, Yan-Li L. and Li, Xiao-Xia X. and Guo, Gang and Zhang, Jing-Yu Y. }, citeulike-article-id = {2770224}, doi = {10.1007/s12033-008-9064-5}, issn = {1073-6085}, journal = {Molecular biotechnology}, keywords = {cloning}, month = {April}, posted-at = {2008-05-08 09:43:36}, priority = {2}, title = {SORS: A Universal One-Round PCR-Based Method for Site-Directed Mutagenesis.}, url = {http://dx.doi.org/10.1007/s12033-008-9064-5}, year = {2008} }

TY - JOUR ID - citeulike:2770224 L3 - citeulike-article-id:2770224 TI - SORS: A Universal One-Round PCR-Based Method for Site-Directed Mutagenesis. JF - Molecular biotechnology AD - Key Research Lab of Hormone and Development Affiliated to the Ministry of Health, Tianjin Medical University, Tianjin, 300070, People’s Republic of China, blwang7212@yahoo.com.cn. SN - 1073-6085 N2 - We have developed a novel protocol for site-directed mutagenesis of double-stranded DNA. The procedure, termed SORS (named because it undergoes the sequential procedure of segmentation-overhang creating PCR-reannealing-splicing) mutagenesis, is exemplified by a substitution, a deletion, and an insertion of nucleotide(s) in target genes. The template DNA is PCR-amplified into two separate segments divided at the prospective mutation site, and each segment is amplified in two parallel PCRs using primers introducing the mutation. The primers are designed to be able to create protruding bases upon pooling, denaturing, and reannealing the two parallel reactions. The protruding bases at the prospective junction of the two segments are mutually complementary; therefore, the two segments can be re-spliced together to generate the mutated gene. Compared to previously published protocols, this procedure is rapid, restriction-independent and ensures higher success rate and lower potential to produce second-site mutations. KW - cloning AU - Wang, Bao-Li AU - Quan, Jin-Xing AU - Liang, Hui AU - Jiao, Yan-Li AU - Li, Xiao-Xia AU - Guo, Gang AU - Zhang, Jing-Yu PY - 2008/04/16/ UR - http://dx.doi.org/10.1007/s12033-008-9064-5 ER -