Lee James Chun-I, Hsieh Hsing-Mei, Teng Hsiao-Feng, Tsai Li-Chin
Department of Forensic Medicine, National Taiwan University, Taipei, Taiwan ROC.
Methods Mol Biol. 2013;919:113-20. doi: 10.1007/978-1-62703-029-8_11.
Single-strand conformation polymorphism by capillary electrophoresis (SSCP-CE) has been developed to detect single nucleotide mutations. This method is used to identify the ABO alleles A(1), A(1v), B, O(1), O(1v), and O(2) in this chapter. Four amplicons (112, 121, 123, and 160 bp) labeled with fluorescence are separately amplified by polymerase chain reaction from exons 6 and 7 of ABO gene. These four fragments are combined into a single tube for SSCP-CE analysis using native gel to identify their single nucleotide polymorphism. This method can fast screen ABO genotypes from unknown samples and will be valuable in clinical transfusion or forensic applications.
毛细管电泳单链构象多态性分析(SSCP-CE)已被开发用于检测单核苷酸突变。本章使用该方法鉴定ABO等位基因A(1)、A(1v)、B、O(1)、O(1v)和O(2)。通过聚合酶链反应从ABO基因的第6和第7外显子分别扩增出四个用荧光标记的扩增子(112、121、123和160 bp)。将这四个片段合并到一个管中,使用天然凝胶进行SSCP-CE分析以鉴定其单核苷酸多态性。该方法可快速从未知样本中筛选ABO基因型,在临床输血或法医应用中具有重要价值。