TITLE

A high-throughput SNP typing system for genome-wide association studies

AUTHOR(S)
Ohnishi, Y.; Tanaka, T.; Ozaki, K.; Yamada, R.; Suzuki, H.; Nakamura, Y.
PUB. DATE
August 2001
SOURCE
Journal of Human Genetics;2001, Vol. 46 Issue 8, p471
SOURCE TYPE
Academic Journal
DOC. TYPE
Article
ABSTRACT
One of the most difficult issues to be solved in genome-wide association studies is to reduce the amount of genomic DNA required for genotyping. Currently available technologies require too large a quantity of genomic DNA to genotype with hundreds or thousands of single-nucleotide polymorphisms (SNPs). To overcome this problem, we combined the Invader assay with multiplex polymerase chain reaction (PCR), carried out in the presence of antibody to Taq polymerase, as well as using a novel 384-well card system that can reduce the required reaction volume. We amplified 100 genomic DNA fragments, each containing one SNP, in a single tube, and analyzed each SNP with the Invader assay. This procedure correctly genotyped 98 of the 100 SNP loci examined in PCR-amplified samples from ten individuals; the genotypes were confirmed by direct sequencing. The reproducibility and universality of the method were confirmed with two additional sets of 100 SNPs. Because we used 40 ng of genomic DNA as a template for multiplex PCR, the amount needed to assay one SNP was only 0.4 ng; therefore, theoretically, more than 200,000 SNPs could be genotyped at once when 100 μg of genomic DNA is available. Our results indicate the feasibility of undertaking genome-wide association studies using blood samples of only 5–10 ml.
ACCESSION #
5352396

 

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