2014年11月
Oligoribonucleotide (ORN) Interference-PCR (ORNi-PCR): A Simple Method for Suppressing PCR Amplification of Specific DNA Sequences Using ORNs
PLOS ONE
- ,
- ,
- 巻
- 9
- 号
- 11
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1371/journal.pone.0113345
- 出版者・発行元
- PUBLIC LIBRARY SCIENCE
Polymerase chain reaction (PCR) amplification of multiple templates using common primers is used in a wide variety of molecular biological techniques. However, abundant templates sometimes obscure the amplification of minor species containing the same primer sequences. To overcome this challenge, we used oligoribonucleotides (ORNs) to inhibit amplification of undesired template sequences without affecting amplification of control sequences lacking complementarity to the ORNs. ORNs were effective at very low concentrations, with IC50 values for ORN-mediated suppression on the order of 10 nM. DNA polymerases that retain 3'-5' exonuclease activity, such as KOD and Pfu polymerases, but not those that retain 5'-3' exonuclease activity, such as Taq polymerase, could be used for ORN-mediated suppression. ORN interference-PCR (ORNi-PCR) technology should be a useful tool for both molecular biology research and clinical diagnosis.
- リンク情報
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- DOI
- https://doi.org/10.1371/journal.pone.0113345
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/25405983
- PubMed Central
- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4236179
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000347121300106&DestApp=WOS_CPL
- URL
- http://europepmc.org/abstract/med/25405983
- URL
- http://orcid.org/0000-0003-1296-4256
- ID情報
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- DOI : 10.1371/journal.pone.0113345
- ISSN : 1932-6203
- ORCIDのPut Code : 49884988
- PubMed ID : 25405983
- PubMed Central 記事ID : PMC4236179
- Web of Science ID : WOS:000347121300106