Reference - Detail
|Title||A novel series of high-efficiency vectors for TA cloning and blunt-end cloning of PCR products.|
An efficient PCR cloning method is indispensable in modern molecular biology, as it can greatly improve the efficiency of DNA cloning processes. Here, I describe the development of three vectors for TA cloning and blunt-end cloning. Specifically, pCRT and pCRZeroT were designed to improve the efficiency of TA cloning. pCRZeroT can also be used with pCRZero to facilitate blunt-end cloning using the ccdB gene. Using pCRZero and pCRZeroT and applying the Golden Gate reaction, I developed a direct PCR cloning protocol with non-digested circular vectors and PCR products. This direct PCR cloning protocol yielded colony-formation rates and cloning efficiencies that are comparable with those obtained by conventional PCR cloning with pre-digested vectors and PCR products. The three plasmids I designed are available from Addgene ( https://www.addgene.org/ ).
|MeSH||Bacterial Proteins / genetics* Bacterial Proteins / metabolism Base Sequence Cloning, Molecular / methods* DNA Primers / chemistry DNA Primers / metabolism Escherichia coli / genetics* Escherichia coli / metabolism Gene Expression Genetic Engineering / methods* Humans Plasmids / chemistry Plasmids / metabolism Polymerase Chain Reaction / methods*|
|DNA material||pCRT (RDB17479) CRZeroT (RDB17480) pCRZero (RDB17481)|