Lallas T A, Buller R E
Department of Obstetrics and Gynecology, University of Iowa Hospitals and Clinics, Iowa City, Iowa 52242, USA.
Mol Genet Metab. 1998 Jul;64(3):173-6. doi: 10.1006/mgme.1998.2713.
Single-strand confirmation polymorphism (SSCP) analysis on MDE gels has been extensively used to detect BRCA1 gene mutations. In screening a large cohort of ovarian cancer patients, our group was not completely satisfied with the standard published techniques. Although conventional primer sets usually amplified well, we were able to enhance band spread and resolution by varying the PCR and electrophoresis conditions for many of the individual exons. These alterations enhanced our ability to detect polymorphisms and mutations. When utilizing SSCP screening of a gene as large as BRCA1, no one set of conditions or even method may be optimal for all of the exons. Rather, a variety of different methods and conditions should be studied for each set of amplimers under analysis.
在MDE凝胶上进行的单链构象多态性（SSCP）分析已被广泛用于检测BRCA1基因突变。在对一大群卵巢癌患者进行筛查时，我们团队对已发表的标准技术并不完全满意。尽管传统引物组通常扩增效果良好，但通过改变许多单个外显子的PCR和电泳条件，我们能够增强条带扩散和分辨率。这些改变提高了我们检测多态性和突变的能力。当利用SSCP对像BRCA1这样大的基因进行筛查时，没有一套条件甚至方法可能对所有外显子都是最佳的。相反，对于每组正在分析的扩增子，应该研究各种不同的方法和条件。
