Higasa Koichiro, Kukita Yoji, Baba Shingo, Hayashi Kenshi
Kyushu University, Fukuoka, Japan.
Biotechniques. 2002 Dec;33(6):1342-8. doi: 10.2144/02336bc05.
PCR single-stranded conformational polymorphism (SSCP) analysis is a simple and rapid electrophoretic technique for the sensitive detection of sequence variants of PCR products. Here we describe a cross-platform program package, quantitative interpretation of SSCP in capillary array (QUISCA), which allows semi-automated quantitative detection of sequence variants separated by multicolor fluorescence-based SSCP electrophoresis using various capillary array apparatus. The program, together with the QUISCAview as a graphical user interface, takes trace data in ASCII format and processes them with three modules: signal denoising/baseline subtraction, color-matrix construction/application, and calibration of peak positions between multiple capillary runs using internal standard peaks. QUISCA is compatible with data from various widely used capillary array sequencers and is suitable not only for finding or typing SNPs in individual DNAs but also for the accurate estimation of the allele frequencies of many SNPs using a pooled DNA strategy. QUISCA can also serve as a versatile core program for various fragment analyses.
聚合酶链反应单链构象多态性（PCR-SSCP）分析是一种简单快速的电泳技术，用于灵敏检测PCR产物的序列变异。在此，我们描述了一个跨平台程序包——毛细管阵列中SSCP的定量解释（QUISCA），它允许使用各种毛细管阵列仪器，通过基于多色荧光的SSCP电泳对分离的序列变异进行半自动定量检测。该程序与作为图形用户界面的QUISCAview一起，获取ASCII格式的痕量数据，并通过三个模块对其进行处理：信号去噪/基线扣除、颜色矩阵构建/应用以及使用内标峰对多个毛细管电泳运行之间的峰位置进行校准。QUISCA与各种广泛使用的毛细管阵列测序仪的数据兼容，不仅适用于在个体DNA中查找或分型单核苷酸多态性（SNP），还适用于使用混合DNA策略准确估计许多SNP的等位基因频率。QUISCA还可作为各种片段分析的通用核心程序。
