Piggee C A, Muth J, Carrilho E, Karger B L
Barnett Institute, Northeastern University, Boston, MA 02115, USA.
J Chromatogr A. 1997 Sep 26;781(1-2):367-75. doi: 10.1016/s0021-9673(97)00637-7.
Capillary electrophoresis (CE) with laser-induced fluorescence (LIF) was used to detect known point mutations using the method of single-nucleotide primer extension (SNuPE). Three different point mutations in human mitochondrial DNA associated with Leber's hereditary optic neuropathy (LHON) were detected by annealing a primer immediately 5' to the mutation on the template and extending the primer by one fluorescently labeled dideoxy terminator complementary to the mutation. By using two or more differently labeled terminators, both the mutant and wild type could be simultaneously detected. The advantages of using CE-LIF for detecting SNuPE reactions include speed and ease of analysis, absence of radioactivity, and potential for automation.
采用激光诱导荧光（LIF）的毛细管电泳（CE）结合单核苷酸引物延伸（SNuPE）方法来检测已知的点突变。通过在模板上紧靠突变位点5'端退火一个引物，并使用与该突变互补的一个荧光标记双脱氧终止子延伸引物，检测了与Leber遗传性视神经病变（LHON）相关的人类线粒体DNA中的三种不同点突变。通过使用两种或更多种不同标记的终止子，可同时检测突变型和野生型。使用CE-LIF检测SNuPE反应的优点包括分析速度快、操作简便、无放射性以及具有自动化潜力。
