Cloning of polymorphisms (COP)

enrichment of polymorphic sequences from complex genomes.

J. Li, F. Wang, V. Zabarovska, Claes R Wahlestedt, E. R. Zabarovsky

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

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.

Original languageEnglish
JournalNucleic Acids Research
Volume28
Issue number2
StatePublished - Jan 15 2000
Externally publishedYes

Fingerprint

Restriction Fragment Length Polymorphisms
Single Nucleotide Polymorphism
Organism Cloning
Uracil-DNA Glycosidase
Genome
Polymerase Chain Reaction
CpG Islands
Streptavidin
Human Genome
Digestion
Clone Cells
Enzymes
Therapeutics
Mung Bean Nuclease

ASJC Scopus subject areas

  • Genetics

Cite this

Cloning of polymorphisms (COP) : enrichment of polymorphic sequences from complex genomes. / Li, J.; Wang, F.; Zabarovska, V.; Wahlestedt, Claes R; Zabarovsky, E. R.

In: Nucleic Acids Research, Vol. 28, No. 2, 15.01.2000.

Research output: Contribution to journalArticle

@article{d8950b00d2514237afdc90033051bcd9,
title = "Cloning of polymorphisms (COP): enrichment of polymorphic sequences from complex genomes.",
abstract = "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.",
author = "J. Li and F. Wang and V. Zabarovska and Wahlestedt, {Claes R} and Zabarovsky, {E. R.}",
year = "2000",
month = "1",
day = "15",
language = "English",
volume = "28",
journal = "Nucleic Acids Research",
issn = "0305-1048",
publisher = "Oxford University Press",
number = "2",

}

TY - JOUR

T1 - Cloning of polymorphisms (COP)

T2 - enrichment of polymorphic sequences from complex genomes.

AU - Li, J.

AU - Wang, F.

AU - Zabarovska, V.

AU - Wahlestedt, Claes R

AU - Zabarovsky, E. R.

PY - 2000/1/15

Y1 - 2000/1/15

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0034651046&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034651046&partnerID=8YFLogxK

M3 - Article

VL - 28

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

IS - 2

ER -