Confronting two-pair primer design for enzyme-free SNP genotyping based on a genetic algorithm.

Confronting two-pair primer design for enzyme-free SNP genotyping based on a genetic algorithm.
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DOI:
10.1186/1471-2105-11-509
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发表时间:
2010-10-13
期刊:
影响因子:
3
通讯作者:
Chang HW
Chang HW
中科院分区:
生物学4区
文献类型:
--
作者:
Yang CH;Cheng YH;Chuang LY;Chang HW

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聚合酶链反应与对抗双对引物(PCR-CTPP)方法通过添加设计的4条引物产生不同长度的等位基因特异性DNA条带,无需进一步的步骤即可通过电泳实现单核苷酸多态性(SNP)基因分型。这是一种省时、经济、简便的SNP基因分型方法。然而,计算可行的CTPP引物仍然是一个挑战。在本研究中,我们提出了一种基于GA(遗传算法)的方法来设计可行的CTPP引物集,以进行可靠的PCR实验。SLC6A4基因在干船坞实验中得到288个snp,表明该算法提供的ctp引物满足大多数引物约束。以SLC6A4基因中的一个SNP rs12449783为例,进行了电泳分型实验,验证了基于ga的设计方法为SNP基因分型提供了可靠的CTPP引物集。基于ga的CTPP引物设计方法为PCR-CTPP的常见引物约束提供了各种形式的估计。GA-CTPP程序是用JAVA实现的,用户友好的输入界面可以在http://bio.kuas.edu.tw/ga-ctpp/上免费获得。
Polymerase chain reaction with confronting two-pair primers (PCR-CTPP) method produces allele-specific DNA bands of different lengths by adding four designed primers and it achieves the single nucleotide polymorphism (SNP) genotyping by electrophoresis without further steps. It is a time- and cost-effective SNP genotyping method that has the advantage of simplicity. However, computation of feasible CTPP primers is still challenging. In this study, we propose a GA (genetic algorithm)-based method to design a feasible CTPP primer set to perform a reliable PCR experiment. The SLC6A4 gene was tested with 288 SNPs for dry dock experiments which indicated that the proposed algorithm provides CTPP primers satisfied most primer constraints. One SNP rs12449783 in the SLC6A4 gene was taken as an example for the genotyping experiments using electrophoresis which validated the GA-based design method as providing reliable CTPP primer sets for SNP genotyping. The GA-based CTPP primer design method provides all forms of estimation for the common primer constraints of PCR-CTPP. The GA-CTPP program is implemented in JAVA and a user-friendly input interface is freely available at http://bio.kuas.edu.tw/ga-ctpp/.
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