Investigating the utility of combining Φ29 whole genome amplification and highly multiplexed single nucleotide polymorphism BeadArray™ genotypingReportar como inadecuado




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BMC Biotechnology

, 4:15

First Online: 27 July 2004Received: 13 April 2004Accepted: 27 July 2004

Abstract

BackgroundSustainable DNA resources and reliable high-throughput genotyping methods are required for large-scale, long-term genetic association studies. In the genetic dissection of common disease it is now recognised that thousands of samples and hundreds of thousands of markers, mostly single nucleotide polymorphisms SNPs, will have to be analysed. In order to achieve these aims, both an ability to boost quantities of archived DNA and to genotype at low costs are highly desirable. We have investigated Φ29 polymerase Multiple Displacement Amplification MDA-generated DNA product MDA product, in combination with highly multiplexed BeadArray™ genotyping technology. As part of a large-scale BeadArray genotyping experiment we made a direct comparison of genotyping data generated from MDA product with that from genomic DNA gDNA templates.

ResultsEighty-six MDA product and the corresponding 86 gDNA samples were genotyped at 345 SNPs and a concordance rate of 98.8% was achieved. The BeadArray sample exclusion rate, blind to sample type, was 10.5% for MDA product compared to 5.8% for gDNA.

ConclusionsWe conclude that the BeadArray technology successfully produces high quality genotyping data from MDA product. The combination of these technologies improves the feasibility and efficiency of mapping common disease susceptibility genes despite limited stocks of gDNA samples.

Electronic supplementary materialThe online version of this article doi:10.1186-1472-6750-4-15 contains supplementary material, which is available to authorized users.

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Autor: Rebecca Pask - Helen E Rance - Bryan J Barratt - Sarah Nutland - Deborah J Smyth - Meera Sebastian - Rebecca CJ Twells

Fuente: https://link.springer.com/article/10.1186/1472-6750-4-15







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