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Nucleic Acids Research, 2000, Vol. 28, No. 2 E1-e1
© 2000 Oxford University Press

Cloning of polymorphisms (COP): enrichment of polymorphic sequences from complex genomes

Jingfeng Li1,2, Fuli Wang2, Veronika Zabarovska2, Claes Wahlestedt1 and Eugene R. Zabarovsky1,2,3,*

1Center for Genomics Research and 2Microbiology and Tumor Biology Center, Karolinska Institute, 171 77 Stockholm, Sweden and 3Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 117 984 Moscow, Russia

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.

* To whom correspondence should be addressed at: CGR and MTC, Karolinska Institute, Box 280, S-171 77, Stockholm, Sweden. Tel: +46 8 728 67 50; Fax: +46 8 31 94 70: Email: eugzab@ki.se


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