Cloning of polymorphisms (COP): enrichment of polymorphic sequences from complex genomes.
Here we describe a new procedure (cloning of polymorphisms, COP) for enrichment of single nucleotide polymorphisms (SNPs) that represent restriction fragment length polymorphisms (RFLPs). COP would be applicable to the isolation of SNPs from particular regions of the genome, e.g. CpG islands, chromosomal bands, YACs or PAC contigs. A combination of digestion with restriction enzymes, treatment with uracil-DNA glycosylase and mung bean nuclease, PCR amplification and purification with streptavidin magnetic beads was used to isolate polymorphic sequences from the genomes of two human samples. After only two cycles of enrichment, 80% of the isolated clones were found to contain RFLPs. A simple method for the PCR detection of these polymorphisms was also developed.
Li
J
J
Center for Genomics Research, Karolinska Institute, 171 77 Stockholm, Sweden.
Wang
F
F
Zabarovska
V
V
Wahlestedt
C
C
Zabarovsky
E R
ER
D003001:Cloning, Molecular / Q000379:methods*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D045647:DNA Glycosylases; D004587:Electrophoresis, Agar Gel; D015894:Genome, Human; D006801:Humans; D009699:N-Glycosyl Hydrolases / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D020641:Polymorphism, Single Nucleotide; D015719:Single-Strand Specific DNA and RNA Endonucleases / Q000378:metabolism; D051981:Uracil-DNA Glycosidase
D004247:DNA; D015719:Single-Strand Specific DNA and RNA Endonucleases; D045647:DNA Glycosylases; D009699:N-Glycosyl Hydrolases; D051981:Uracil-DNA Glycosidase