Baba S, Kukita Y, Higasa K, Tahira T, Hayashi K
Kyushu University, Fukuoka, Japan.
Biotechniques. 2003 Apr;34(4):746-50. doi: 10.2144/03344st03.
We describe a new environment of a single-stranded conformational polymorphism (SSCP) analysis using automated capillary array sequencers (e.g., ABI Prism 3100 and 3700). In this environment, electrophoretic conditions, settings for instrument management, and software for data analysis are adjusted for SSCP analysis. Highly reproducible results are obtained with this new system, and fragments with mutations and/or polymorphisms in different capillaries or different runs can be reliably detected. The relative peak heights between alleles are quantitative and reproducible between runs, and so allele frequencies of single nucleotide polymorphisms can be accurately estimated by a pooled DNA strategy. The method allows unattended, low-cost, and quantitative SSCP analysis using instruments that are widely accessible.
我们描述了一种使用自动毛细管阵列测序仪（例如ABI Prism 3100和3700）进行单链构象多态性（SSCP）分析的新环境。在这种环境下，针对SSCP分析对电泳条件、仪器管理设置和数据分析软件进行了调整。使用这个新系统可获得高度可重复的结果，并且能够可靠地检测出不同毛细管或不同运行中存在突变和/或多态性的片段。等位基因之间的相对峰高是定量的且在各次运行之间具有可重复性，因此通过混合DNA策略可以准确估计单核苷酸多态性的等位基因频率。该方法允许使用广泛可得的仪器进行无人值守、低成本的定量SSCP分析。
