Restriction primers as short as 6-mers for PCR amplification of bacterial and plant genomic DNA and plant viral RNA
Identifieur interne : 003701 ( Main/Exploration ); précédent : 003700; suivant : 003702Restriction primers as short as 6-mers for PCR amplification of bacterial and plant genomic DNA and plant viral RNA
Auteurs : Ki Hyun Ryu [Corée du Sud] ; Sun Hee Choi ; Jong Suk LeeSource :
- Molecular Biotechnology [ 1073-6085 ] ; 2000-01-01.
Descripteurs français
- KwdFr :
- MESH :
English descriptors
- KwdEn :
- MESH :
- chemical , genetics : DNA Primers, DNA, Bacterial, DNA, Plant, RNA, Viral.
- genetics : Plants, Pseudomonas.
- methods : Polymerase Chain Reaction.
Abstract
Abstract: Amplification of DNA or RNA sequences using the polymerase chain reaction (PCR) or reverse transcriptase PCR (RT-PCR) requires primers of an appropriate length to be designed. Two hexamer restriction primers, denoted as E101 and H301, which correspond to sequences of EcoRI and HindIII recognition sites, respectively, were selected and used as primers in PCR and RT-PCR. We first applied the restriction primers to the plasmid DNA and bacterial (Pseudomonas) and plant (Cymbidium) genomic DNAs. We observed positive DNA amplifications with the recombinant plasmid DNA and bacterial and plant genomic DNAs. Purified viral RNA was used for template in the RT-PCR with the primers and successful DNA amplification was obtained. These results suggest that the 6-mer restriction primers can be useful for new applications in PCR.
Url:
DOI: 10.1385/MB:14:1:01
Affiliations:
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Le document en format XML
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<term>Polymerase Chain Reaction (methods)</term>
<term>Primer</term>
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<term>amplification</term>
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<term>RNA, Viral</term>
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<term>Amorces ADN</term>
<term>Plantes</term>
<term>Pseudomonas</term>
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<front><div type="abstract" xml:lang="en">Abstract: Amplification of DNA or RNA sequences using the polymerase chain reaction (PCR) or reverse transcriptase PCR (RT-PCR) requires primers of an appropriate length to be designed. Two hexamer restriction primers, denoted as E101 and H301, which correspond to sequences of EcoRI and HindIII recognition sites, respectively, were selected and used as primers in PCR and RT-PCR. We first applied the restriction primers to the plasmid DNA and bacterial (Pseudomonas) and plant (Cymbidium) genomic DNAs. We observed positive DNA amplifications with the recombinant plasmid DNA and bacterial and plant genomic DNAs. Purified viral RNA was used for template in the RT-PCR with the primers and successful DNA amplification was obtained. These results suggest that the 6-mer restriction primers can be useful for new applications in PCR.</div>
</front>
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