文献_DOP-PCR1992.pdfVIP

GENOMICS 13, 718-725 (1992) Degenerate Oligonucleotide-Primed PCR: General Amplification of Target DNA by a Single Degenerate Primer HAKAN TELENIUS,*+’ NIGEL P. CARTER, t CHARLOTTE E. BEBB, t MAGNUS NORDENSKJ~LD, $ BRUCE A. J. PONDER,* AND ALAN TUNNACLIFFE” *CRC Human Cancer Genetics Research Group and tHuman Molecular Genetics Group, Department of Pathology, University of Cambridge, United Kingdom; and *Department of Clinical Genetics, Karolinska Hospital, Stockholm, Sweden Received December 12, 1991; revised February 28, 1992 spersed repetitive sequence PCR (IRS-PCR) has in this A version of the polymerase chain reaction (PCR), manner been used to create human chromosome- and termed degenerate oligonucleotide-primed PCR (DOP- region-specific libraries (Nelson et al, 1989; Cotter et al., PCR), which employs oligonucleotides of partially de- 1990, 1991; Ledbetter et al., 1990b). In man, the most generate sequence, has been developed for genome abundant family of repeats is the Ah family, estimated mapping studies. This degeneracy, together with a to comprise 900,000 elements in the haploid genome, PCR protocol utilizing a low initial annealing tempera- thus giving an