Single nucleotide polymorphism genotyping by mini-primer allele-specific amplification with universal reporter primers for identification of degraded DNA
Single nucleotide polymorphism genotyping by mini-primer allele-specific amplification with universal reporter primers for identification of degraded DNA
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DOI：
10.1016/j.ab.2008.11.023
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发表时间：
2009-03-01
影响因子：
2.9
通讯作者：
Shimizu, Keiko
中科院分区：
文献类型：
--
作者：
Asari, Masaru;Watanabe, Satoshi;Shimizu, Keiko
单核苷酸多态性（SNP）为人类鉴定提供了信息，与高度多态性的短串联重复序列（STR）相比，双等位基因SNP分析的目标区域要短得多。因此，SNP基因分型比STR基因分型对降解的人类DNA更敏感。为了实现简单、经济、灵敏的SNP基因分型技术，我们开发了一种基于mini-引物等位基因特异性扩增（ASA）和通用报告引物（URP）的18个位点的SNP基因分型技术。基于URP/ asa的基因分型包括两次扩增和毛细管电泳检测。目标基因组片段长度在40 ~ 67 bp之间。在日本人群中，18个snp的次要等位基因的频率在0.36 ~ 0.50之间，这些snp为鉴定提供了信息。SNP基因分型的成功率远高于人工降解DNA的STR基因分型。此外，我们将这种基因分型方法应用于病例样本，并成功地对来自4年缓冲福尔马林固定组织样本的严重降解DNA进行了SNP基因分型，用于人类鉴定。(c) 2008爱思唯尔公司版权所有。
Single nucleotide polymorphism (SNP) is informative for human identification, and much shorter regions are targeted in analysis of biallelic SNP compared with highly polymorphic short tandem repeat (STR). Therefore, SNP genotyping is expected to be more sensitive than STR genotyping of degraded human DNA. To achieve simple, economical, and sensitive SNP genotyping for identification of degraded human DNA, we developed 18 loci for a SNP genotyping technique based on the mini-primer allele-specific amplification (ASA) combined With universal reporter primers (URP). The URP/ASA-based genotyping consisted of two amplifications followed by detection using capillary electrophoresis. The sizes of the target genome fragments ranged from 40 to 67 bp in length. In the Japanese Population, the frequencies of minor alleles of 18 SNPs ranged from 0.36 to 0.50, and these SNPs are informative for identification. The success rate of SNP genotyping was much higher than that of STR genotyping of artificially degraded DNA. Moreover, we applied this genotyping method to case samples and showed successful SNP genotyping of severely degraded DNA from a 4-year buffered formalin-fixed tissue sample for human identification. (c) 2008 Elsevier Inc. All rights reserved.