Reference - Detail
RRC ID | 57636 |
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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 | 3.998 |
Times Cited | 3 |
Resource | |
DNA material | pCRT (RDB17479) CRZeroT (RDB17480) pCRZero (RDB17481) |