論文 - 詳細
| RRC ID | 57636 |
|---|---|
| 著者 | Motohashi K. |
| タイトル | A novel series of high-efficiency vectors for TA cloning and blunt-end cloning of PCR products. |
| ジャーナル | 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/ ). |
| 巻・号 | 9(1) |
| ページ | 6417 |
| 公開日 | 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 |
| 引用数 | 3 |
| リソース情報 | |
| 遺伝子材料 | pCRT (RDB17479) CRZeroT (RDB17480) pCRZero (RDB17481) |