本文選題：單核苷酸多態(tài)性(SNP) + 二代測(cè)序　； 參考：《東華大學(xué)學(xué)報(bào)(自然科學(xué)版)》2017年03期

【摘要】：通過(guò)將二代測(cè)序數(shù)據(jù)與連接酶檢測(cè)反應(yīng)(ligase detection reaction,LDR)對(duì)單核苷酸多態(tài)性(single nucleotide polymorphism,SNP)基因分型的結(jié)果進(jìn)行比對(duì),確定二代測(cè)序數(shù)據(jù)判定SNP基因型的經(jīng)驗(yàn)性閾值.利用多重聚合酶鏈?zhǔn)椒磻?yīng)(multiplex polymerase chain reaction,multiplex PCR)對(duì)91個(gè)樣本進(jìn)行19個(gè)SNP位點(diǎn)的擴(kuò)增,擴(kuò)增的產(chǎn)物混勻純化后在Ion torrent PGM儀器上進(jìn)行二代測(cè)序.利用LDR技術(shù)對(duì)相應(yīng)的SNP位點(diǎn)進(jìn)行檢測(cè),將其分型結(jié)果作為二代測(cè)序數(shù)據(jù)判定SNP基因型的標(biāo)準(zhǔn),確定了二代測(cè)序數(shù)據(jù)判定SNP基因型的閾值:測(cè)序深度≥6X,等位基因比率在15%~85%的位點(diǎn)為雜合子,在范圍之外的為純合子,該閾值準(zhǔn)確度達(dá)到99.6%;針對(duì)等位基因頻率分布在閾值邊緣的數(shù)據(jù),結(jié)合聚類分析可將正確率提升至100%.研究結(jié)果為利用二代測(cè)序數(shù)據(jù)判定SNP基因型提供了一個(gè)準(zhǔn)確、快捷和經(jīng)驗(yàn)性的閾值與方案.
[Abstract]:The results of single nucleotide polymorphisms single nucleotide polymorphismSNPs genotyping were compared with ligase detection reactionation (LDRs) of the second generation sequencing data, and the empirical threshold for determining the genotypes of SNPs was determined by sequencing data of the second generation. Nineteen SNPs of 91 samples were amplified by multiplex polymerase chain reaction multiplex PCR. The amplified products were mixed and purified and sequenced by second generation on Ion torrent PGM instrument. The corresponding SNP loci were detected by using LDR technique, and the typing results were used as the criteria for judging the SNP genotypes by the second-generation sequencing data. The threshold for determining SNP genotypes by second-generation sequencing data was determined: the sequencing depth 鈮，

本文編號(hào)：2044103