In vitro amplification of whole large plasmids via transposon-mediated oriC insertion
DNA amplification is a fundamental technique in molecular biology. The replication cycle reaction is a new method for amplification of large circular DNA having oriC sequences, which is a replication initiation site of the Escherichia coli chromosome. We here developed a replication cycle reaction-b...
| Veröffentlicht in: | BioTechniques. - 1988. - 71(2021), 4 vom: 31. Okt., Seite 528-533 |
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| Format: | Online-Aufsatz |
| Sprache: | English |
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2021
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| Zugriff auf das übergeordnete Werk: | BioTechniques |
| Schlagworte: | Journal Article Research Support, Non-U.S. Gov't DNA amplification circular DNA plasmid replication cycle reaction transposon DNA, Bacterial DNA, Circular |
| Zusammenfassung: | DNA amplification is a fundamental technique in molecular biology. The replication cycle reaction is a new method for amplification of large circular DNA having oriC sequences, which is a replication initiation site of the Escherichia coli chromosome. We here developed a replication cycle reaction-based method useful for amplification of various circular DNAs lacking oriC, even in the absence of any sequence information, via transposon-mediated oriC insertion to the circular DNA template. A 15-kb non-oriC plasmid was amplified from a very small amount of starting DNA (50 fg, 1 fM). The method was also applicable to GC-rich plasmid (69%) or large F-plasmid (230 kb). This method thus provides a powerful tool to amplify various environmental circular DNAs |
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| Beschreibung: | Date Completed 06.12.2021 Date Revised 14.12.2021 published: Print-Electronic Citation Status MEDLINE |
| ISSN: | 1940-9818 |
| DOI: | 10.2144/btn-2021-0068 |