Genotypic analysis of multiple loci in somatic cells by whole genome amplification

Michael T. Barrett; Brian J. Reid; Geoffrey Joslyn

doi:10.1093/nar/23.17.3488

Genotypic analysis of multiple loci in somatic cells by whole genome amplification

Michael T. Barrett, Brian J. Reid, Geoffrey Joslyn

Health Sciences Research

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Abstract

To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Aliquots of these reactions were then used in locus-specific reactions using a single round of PCR cycles with individual sets of primers representing polymorphic markers for different regions. As a result, polymorphic markers for different chromosomal regions, including VNTRs and dinucleotide repeats, can be used to perform up to 30 locus-specific PCR assays with a single sample obtained from fewer than 1000 cells.

Original language	English (US)
Pages (from-to)	3488-3492
Number of pages	5
Journal	Nucleic acids research
Volume	23
Issue number	17
DOIs	https://doi.org/10.1093/nar/23.17.3488
State	Published - Sep 11 1995

ASJC Scopus subject areas

Genetics

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title = "Genotypic analysis of multiple loci in somatic cells by whole genome amplification",

abstract = "To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Aliquots of these reactions were then used in locus-specific reactions using a single round of PCR cycles with individual sets of primers representing polymorphic markers for different regions. As a result, polymorphic markers for different chromosomal regions, including VNTRs and dinucleotide repeats, can be used to perform up to 30 locus-specific PCR assays with a single sample obtained from fewer than 1000 cells.",

author = "Barrett, {Michael T.} and Reid, {Brian J.} and Geoffrey Joslyn",

note = "Funding Information: We gratefully acknowledge James K. McDougall for his continued encouragement and support of this work and Carissa A. Sanchez for DNA content cell sorting. This work was supported by NIH grant CA61202, NIH grant CA42792 and American Cancer Society grant EDT-21E.",

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AU - Barrett, Michael T.

AU - Reid, Brian J.

AU - Joslyn, Geoffrey

N1 - Funding Information: We gratefully acknowledge James K. McDougall for his continued encouragement and support of this work and Carissa A. Sanchez for DNA content cell sorting. This work was supported by NIH grant CA61202, NIH grant CA42792 and American Cancer Society grant EDT-21E.

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Y1 - 1995/9/11

N2 - To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Aliquots of these reactions were then used in locus-specific reactions using a single round of PCR cycles with individual sets of primers representing polymorphic markers for different regions. As a result, polymorphic markers for different chromosomal regions, including VNTRs and dinucleotide repeats, can be used to perform up to 30 locus-specific PCR assays with a single sample obtained from fewer than 1000 cells.

AB - To screen multiple loci in small purified samples of diploid and aneuploid cells a PCR-based technique of whole genome amplification was adapted to the study of somatic lesions. DNA samples from different numbers of flow-sorted diploid and aneuploid cells from biopsies were amplified with a degenerate 15mer primer. Aliquots of these reactions were then used in locus-specific reactions using a single round of PCR cycles with individual sets of primers representing polymorphic markers for different regions. As a result, polymorphic markers for different chromosomal regions, including VNTRs and dinucleotide repeats, can be used to perform up to 30 locus-specific PCR assays with a single sample obtained from fewer than 1000 cells.

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Genotypic analysis of multiple loci in somatic cells by whole genome amplification

Abstract

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