Degenerate oligonucleotide-primed PCR: general amplification of target DNA by a single degenerate primer.
A version of the polymerase chain reaction (PCR), termed degenerate oligonucleotide-primed PCR (DOP-PCR), which employs oligonucleotides of partially degenerate sequence, has been developed for genome mapping studies. This degeneracy, together with a PCR protocol utilizing a low initial annealing temperature, ensures priming from multiple (e.g., approximately 10(6) in human) evenly dispersed sites within a given genome. Furthermore, as efficient amplification is achieved from the genomes of all species tested using the same primer, the method appears to be species-independent. Thus, for the general amplification of target DNA, DOP-PCR has advantages over interspersed repetitive sequence PCR (IRS-PCR), which relies on the appropriate positioning of species-specific repeat elements. In conjunction with chromosome flow sorting, DOP-PCR has been applied to the characterization of abnormal chromosomes and also to the cloning of new markers for specific chromosome regions. DOP-PCR therefore represents a rapid, efficient, and species-independent technique for general DNA amplification.
Telenius
H
H
Department of Pathology, University of Cambridge, United Kingdom.
Carter
N P
NP
Bebb
C E
CE
Nordenskjöld
M
M
Ponder
B A
BA
Tunnacliffe
A
A
D000818:Animals; D001483:Base Sequence; D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004247:DNA / Q000235:genetics*; D005069:Evaluation Studies as Topic; D005819:Genetic Markers; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D009838:Oligodeoxyribonucleotides / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*
D005819:Genetic Markers; D009838:Oligodeoxyribonucleotides; D004247:DNA