论文部分内容阅读
目的 :建立检测MTHFRC6 77T点突变的PCR -TGGE技术 .方法 :以中国人群外周血基因组DNA为模板 ,用聚合酶链反应扩增该基因外显子 4 ,用温度梯度凝胶电泳 (TGGE)分析检测PCR产物 ,对电泳结果异常者进行DNA测序 .结果 :垂直型TGGE与平行型TGGE均发现该外显子存在杂合性突变 ,经DNA测序证实为C6 77T点突变 .结论 :该技术具有快速、分辨能力高和重复性好的特点 ,是一种有效的检测核酸序列变异和点突变的技术 .
OBJECTIVE: To establish a PCR-TGGE technique for detection of MTHFRC6 77T point mutation.Methods: Exon 4 of this gene was amplified by polymerase chain reaction using genomic DNA from Chinese population as a template and analyzed by temperature gradient gel electrophoresis (TGGE) The PCR products were detected and DNA sequencing was performed on those with abnormal electrophoresis results.Results: The heterozygous mutations were found in both vertical TGGE and parallel TGGE, and C677T point mutation was confirmed by DNA sequencing.Conclusion: This technique has the advantages of fast , High resolving power and good repeatability, it is an effective technique to detect nucleic acid sequence variation and point mutation.