Single-nucleotide polymorphism analysis by pyrosequencing
Single-nucleotide polymorphism analysis by pyrosequencing
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DOI：
10.1006/abio.2000.4493
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发表时间：
2000-04-10
影响因子：
2.9
通讯作者：
Lundeberg, J
中科院分区：
文献类型：
--
作者：
Ahmadian, A;Gharizadeh, B;Lundeberg, J
对于分析单核苷酸多态性（SNP）位置的高通量方法的需求不断增长。在这里，出于此目的，我们评估了一种新型的测序方法，即焦磷酸测序。焦磷酸测序是一种逐一测序方法，其中一系列酶促反应产生可检测的光，与掺入核苷酸成正比。与其他两个变体相比，用焦磷酸测序键入SNP的一个功能是，每个等位基因变体都将提供一个独特的序列。这些变体很容易通过模式识别软件来区分。该软件显示等位基因：替代方案，并允许与pyrosequencing原始数据进行直接比较。为了获得SNP的最佳测定，研究了各种核苷酸分配顺序的方案。在这里，我们证明了SNP的键入可以通过使用自动化系统在大约5分钟内使用自动化系统进行焦磷酸测序进行并行分析，适用于SNP的大规模筛选和分型。 （c）2000学术出版社。
There is a growing demand for high-throughput methods for analysis of single-nucleotide polymorphic (SNP) positions. Here, we have evaluated a novel sequencing approach, pyrosequencing, for such purposes. Pyrosequencing is a sequencing-by-synthesis method in which a cascade of enzymatic reactions yields detectable light, which is proportional to incorporated nucleotides. One feature of typing SNPs with pyrosequencing is that each allelic variant will give a unique sequence compared to the two other variants. These variants can easily be distinguished by a pattern recognition software. The software displays the allelic: alternatives and allows for direct comparison with the pyrosequencing raw data. For optimal determination of SNPs, various protocols of nucleotide dispensing order were investigated. Here, we demonstrate that typing of SNPs can efficiently be performed by pyrosequencing using an automated system for parallel analysis of 96 samples in approximately 5 min, suitable for large-scale screening and typing of SNPs. (C) 2000 Academic Press.