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The use of whole genome amplification in the study of human disease
Authors:Hughes Simon  Arneson Nona  Done Susan  Squire Jeremy
Affiliation:Ontario Cancer Institute, Princess Margaret Hospital, University Health Network, Ontario, Canada. sih3@yahoo.com
Abstract:The availability of large amounts of genomic DNA is of critical importance for many of the molecular biology assays used in the analysis of human disease. However, since the amount of patient tissue available is often limited and as particular foci of interest may consist of only a few hundred cells, the yield of DNA is often insufficient for extensive analysis. To address this problem, several whole genome amplification (WGA) methodologies have been developed. Initial WGA approaches were based on the polymerase chain reaction (PCR). However, recent reports have described the use of non-PCR-based linear amplification protocols for WGA. Using these methods, it is possible to generate microgram quantities of DNA starting with as little as 1mg of genomic DNA. This review will provide an overview of WGA approaches and summarize some of the uses for amplified DNA in various high-throughput genetic applications.
Keywords:WGA, whole genome amplification   PCR, polymerase chain reaction   PEP, primer extension preamplification   DOP-PCR, degenerate oligonucleotide primed PCR   LMP, ligation mediated PCR   SDA, strand displacement amplification   TLAD, T7-based linear amplification of DNA   LCM, laser capture microdissection   IRS, interspersed repetitive sequence   LA-PCR, linker adapter technique PCR   LL-DOP-PCR, long products from low DNA quantities DOP-PCR   I-PEP, improved PEP   CGH, comparative genomic hybridization   SNP, single nucleotide polymorphism   SSCP, single stranded conformational polymorphism   RFLP, restriction fragment length polymorphism   EFPE, ethanol-fixed paraffin embedded   FFPE, formalin-fixed paraffin embedded   LOH, loss of heterozygosity   TSG, tumour suppressor genes   SCOMP, single cell comparative genomic hybridization   PRSG, PCR of randomly sheared genomic   MDA, multiple displacement amplification   RCA, rolling circle amplification
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