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Comparison of whole genome amplification methods for further quantitative analysis with microarray-based comparative genomic hybridization.

Abstract：

OBJECTIVE:Whole genome amplification (WGA) is a crucial procedure for genomic DNA analysis from limited sources, such as in forensic analysis, embryo biopsy for preimplantation genetic diagnosis, or needle aspiration biopsies. Several strategies for WGA have been developed for either genotyping or microarray-based comparative genome hybridization (array-CGH) during the past decade. Nevertheless, there were few studies in which various WGA methods had been performed side-by-side and results evaluated with multiple methods. MATERIALS AND METHODS:Ease of performance, qualitative accuracy, and quantitative fidelity of different WGA methods, such as degenerate oligonucleotide-primed polymerase chain reaction (DOP-PCR), ligation-mediated PCR (LM-PCR) and strand displacement amplification (SDA), were compared in amplifying genomic DNA derived from karyotype-confirmed amniocytes and the cancer cell line SAOS2. RESULTS:Using analysis with microsatellite markers, single nucleotide polymorphism markers, and array-CGH, our results suggested that: (1) genomic DNA amplified from DOP-PCR resulted in false positive and negative results by analysis with array-CGH; (2) SDA is the easiest performance method among the three WGA methods; and (3) amplified DNA products generated by LM-PCR best reflect the original genomic DNA. CONCLUSION:The amplified DNA products generated by LM-PCR best reflect the original genomic DNA.

journal_name

Taiwan J Obstet Gynecol

journal_title

Taiwanese journal of obstetrics & gynecology

authors

Lee YS,Tsai CN,Tsai CL,Chang SD,Hsueh DW,Liu CT,Ma CC,Lin SH,Wang TH,Wang HS

doi

10.1016/S1028-4559(08)60052-2

subject

Has Abstract

pub_date

2008-03-01 00:00:00

pages

32-41

issue

1

eissn

1028-4559

issn

1875-6263

pii

S1028-4559(08)60052-2

journal_volume

47

pub_type

杂志文章

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