Phelps R S, Chadwick R B, Conrad M P, Kronick M N, Kamb A
Myriad Genetics, Inc., Salt Lake City, UT, USA.
Biotechniques. 1995 Dec;19(6):984-9.
A crucial factor in the success of positional cloning efforts is the ability to screen rapidly many different candidate genes for mutations. By modifying standard software, we have improved the detection of heterozygous base positions in PCR products sequenced by cycle sequencing. A key element of the method is the incorporation of a modified heterozygote detection algorithm that permits the use of DNA sequence data derived from PCR and sequencing reactions that have not been fully optimized. This allows sequencing runs of average quality to be used. We demonstrate that the sensitivity and specificity of the method are well suited to mutation detection applications such as positional cloning.
定位克隆研究成功的一个关键因素是能够快速筛选许多不同的候选基因突变。通过修改标准软件，我们改进了对循环测序法测定的PCR产物中杂合碱基位置的检测。该方法的一个关键要素是采用了一种改进的杂合子检测算法，该算法允许使用来自尚未完全优化的PCR和测序反应的DNA序列数据。这使得平均质量的测序运行也能得以利用。我们证明，该方法的灵敏度和特异性非常适合于定位克隆等突变检测应用。
