Establishment and application of a method for high-throughput screening single nucleotide polymorphism by tag microarray / 解放军医学杂志

ABSTRACT

Objective To establish a method for high throughput screening single nucleotide polymorphism(SNP) by tag microarray,and then apply the method to study the gene SNP which is related to the motor function of normal people.Methods The genes related to motor function were firstly defined,and then 48 SNP loci were determined.The rs numbers of these SNP loci were fingered out from PubMed,and the primers were designed with the software in web site "www.autoprimer.com".The primer sequences were then downloaded and sent to the biologic corporation for synthesis.After being synthesized and purified by HPLC these primers were used in the experiments according to the instruction of Bakeman's SNPstream machine.The key techniques of SNPstream machine were tag microarray and single nucleotide extension assay.Once the determination was finished,both the gene frequency and allele frequency of every locus could be statistically analyzed.Results The information of the 48 SNP loci that related to motor function had been determined simultaneously by tag microarray,regardless the number of samples to be detected at the same time.The number of the samples was variable to meet the need.The data of gene frequency and allele frequency of these 48 SNP loci may be used in the subsequent studies.Conclusions Tag microarray used to high throughput screening SNP has the advantages of accuracy,speed,efficiency and reasonable cost.Therefore it can be applied to study the relationship between the SNP and many kinds of diseases.