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The use of whole genome amplification in the study of human disease |
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| Authors: | Hughes Simon Arneson Nona Done Susan Squire Jeremy |
| Institution: | 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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