Supported PCR: an efficient procedure to amplify sequences flanking a known DNA segment

George N. Rudenko, Caius M.T. Rommens, H. John J. Nijkamp, Jacques Hille

OnderzoeksoutputAcademic

10 Citaten (Scopus)
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Samenvatting

We describe a novel modification of the polymerase chain reaction for efficient in vitro amplification of genomic DNA sequences flanking short stretches of known sequence. The technique utilizes a target enrichment step, based on the selective isolation of biotinylated fragments from the bulk of genomic DNA on streptavidin-containing support. Subsequently, following ligation with a second universal linker primer, the selected fragments can be amplified to amounts suitable for further molecular studies. The procedure has been applied to recover T-DNA flanking sequences in transgenic tomato plants which could subsequently be used to assign the positions of T-DNA to the molecular map of tomato. The method called supported PCR (sPCR) is a simple and efficient alternative to techniques used in the isolation of specific sequences flanking a known DNA segment.
Originele taal-2English
Aantal pagina's6
TijdschriftPlant Molecular Biology
Volume21
Nummer van het tijdschrift4
DOI's
StatusPublished - 1993

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