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Plant Mol Biol. 1993 Feb;21(4):723-8.

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

Author information

1
Free University, Department of Genetics, Amsterdam, Netherlands.

Abstract

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.

PMID:
8448372
DOI:
10.1007/bf00014557
[Indexed for MEDLINE]

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