SYBR green dye-based probe-free SNP genotyping: Introduction of T-Plex real-time PCR assay
SYBR green dye-based probe-free SNP genotyping: Introduction of T-Plex real-time PCR assay
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DOI：
10.1016/j.ab.2013.07.007
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发表时间：
2013-10-15
影响因子：
2.9
通讯作者：
Baris, Saniye Tugba
中科院分区：
文献类型：
--
作者：
Baris, Ibrahim;Etlik, Ozdal;Baris, Saniye Tugba
单核苷酸多态性（SNP）基因分型被广泛用于遗传关联研究，以表征遗传性状背后的遗传因素。尽管最近在高通量SNP基因分型方面取得了许多进展，但仍然需要具有合理通量水平的廉价且灵活的方法。用于发现和基因分型SNP的实时PCR方法在生物学的各个领域中变得越来越重要。在这项研究中，我们介绍了一种新的，单管策略，结合四引物ARMS PCR检测，SYBR绿色I为基础的实时PCR，熔点分析与引物设计策略，以检测感兴趣的SNP。T-PCR实时荧光定量PCR是基于T.在单个管中区分扩增的等位基因特异性扩增子。针对FV、PII、MTHFR和FGFR 3基因中的常见突变，评价了该检测方法的特异性、灵敏度和耐用性。我们相信，T-PCR实时PCR将是一个有用的替代方案，无论是个人的基因分型要求或大型流行病学研究。(C)2013 Elsevier Inc. All rights reserved.
Single-nucleotide polymorphism (SNP) genotyping is widely used in genetic association studies to characterize genetic factors underlying inherited traits. Despite many recent advances in high-throughput SNP genotyping, inexpensive and flexible methods with reasonable throughput levels are still needed. Real-time PCR methods for discovering and genotyping SNPs are becoming increasingly important in various fields of biology. In this study, we introduce a new, single-tube strategy that combines the tetra-primer ARMS PCR assay, SYBR Green I-based real-time PCR, and melting-point analysis with primer design strategies to detect the SNP of interest. This assay, T-Plex real-time PCR, is based on the T. discrimination of the amplified allele-specific amplicons in a single tube. The specificity, sensitivity, and robustness of the assay were evaluated for common mutations in the FV, PII, MTHFR, and FGFR3 genes. We believe that T-Plex real-time PCR would be a useful alternative for either individual genotyping requests or large epidemiological studies. (C) 2013 Elsevier Inc. All rights reserved.