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Tag/anti-tag liquid-phase primer extension array: a flexible and versatile genotyping platform.

机译:标签/反标签液相引物延伸阵列:灵活而通用的基因分型平台。

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摘要

This study demonstrates an array-based platform to genotype simultaneously single nucleotide polymorphisms (SNPs) and some short insertions/deletions (indels) by the integration of the universal tag/anti-tag (TAT) system, liquid-phase primer extension (LIPEX), and a novel two-color detection strategy on an array format (TATLIPEXA). The TAT system permits a universal chip to be used for many applications, and the LIPEX simplifies the sample preparation but improves the sensitivity significantly. More importantly, all SNPs and some short indels can be interrogated in a single reaction with only two fluorescent ddNTPs. The concept of TATLIPEXA is demonstrated for nine SNPs (eight point mutations and one single-base insertion), and genotypes obtained show a remarkable concordance rate of 100% with both DNA sequencing and restriction fragment length polymorphism. Moreover, TATLIPEXA is able to provide quantitative information on allele frequency in pooled DNA samples, which could serve as a rapid screening tool for SNPs associated with diseases.
机译:这项研究展示了一个基于阵列的平台,通过整合通用标签/抗标签(TAT)系统,液相引物延伸(LIPEX),可以同时对单核苷酸多态性(SNP)和一些短插入/缺失(indels)进行基因分型。 ,以及基于数组格式(TATLIPEXA)的新颖的两色检测策略。 TAT系统允许通用芯片用于许多应用,而LIPEX简化了样品制备,但显着提高了灵敏度。更重要的是,仅使用两个荧光ddNTP即可在单个反应中询问所有SNP和一些短indel。证明了TATLIPEXA的概念可用于9个SNP(8个点突变和1个单碱基插入),获得的基因型与DNA测序和限制性片段长度多态性均显示出100%的显着一致性。此外,TATLIPEXA能够提供有关合并DNA样本中等位基因频率的定量信息,这可以用作与疾病相关的SNP的快速筛选工具。

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