RRC ID 57636
Author Motohashi K.
Title A novel series of high-efficiency vectors for TA cloning and blunt-end cloning of PCR products.
Journal Sci Rep
Abstract 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/ ).
Volume 9(1)
Pages 6417
Published 2019-4-23
DOI 10.1038/s41598-019-42868-6
PII 10.1038/s41598-019-42868-6
PMID 31015513
PMC PMC6478821
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*
IF 4.011
Times Cited 3
Resource
DNA material pCRT (RDB17479) CRZeroT (RDB17480) pCRZero (RDB17481)