贵州医科大学学报

2006, (01) 18-20

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直接测序法与克隆测序法在单核苷酸多态性检测中的比较
A Comparison of Direct Sequencing with Clone Sequencing in the Detection of Single Nucleotide Polymorphism

张婷;单可人;李毅;何燕;齐晓岚;赵艳;谢渊;吴昌学;任锡麟;官志忠;
ZHANG Ting,SHAN Ke-ren,LI Yi,HE Yan,QI Xiao-lan,ZHAO Yan,XIE Yuan,Wu Chang-xue,REN Xi-lin,GUAN Zhi-zhong(Molecular Biological Key Laboratory of Guiyang Medical College,Guiyang 550004,China)

摘要(Abstract):

目的:了解PCR产物直接测序法与克隆测序法在单核苷酸多态性(SNP)检测上的差别。方法:对2型糖尿病患者的载脂蛋白J外显子2基因及旁侧进行聚合酶链反应(PCR)扩增,分别对其产物进行直接测序和克隆测序。结果:在样本的检测中,PCR产物直接测序法所测序列与43~60 bp以后克隆测序相同;检出一个突变位点并与GenBank(DQ012938)发表序列一致。结论:PCR产物直接测序法和克隆测序法都是检测SNP的有效方法,但前者不仅特异性强,敏感度高,而且比克隆测序方法更为快速简便,节省材料。
Objective: To compare the sensitivity and accuracy of the 2 sequencing techniques for the detection of single nucleotide polymorphism(SNP).Methods: The exon 2 of Apo J gene was screened with polymerase chain reaction(PCR)in 8 patients with type 2 diabetes mellitus(DM).Samples were detected by direct sequencing and clone sequencing,and the results were compared.Results: One mutation was detected in exon 2 by two methods.Furthermore,it's same to the Homo sapiens apolipoprotein J(CLU) gene sequence in GenBank(DQ012938).Conclusion: The results show that the PCR direct sequencing is faster,easier,cheaper,and more practical than clone sequencing when used in SNP.

关键词(KeyWords): 载脂蛋白类;多态性,单核苷酸;糖尿病,非胰岛素依赖型;聚合酶链反应;直接测序法;克隆测序法
apolipoprateins;polymorphism,single nucleotide;diabetes mellitus,non-insulin-dependent;polymerase chain reaction;direct sequencing;clone sequencing

Abstract:

Keywords:

基金项目(Foundation): 贵州省科技厅科研基金资助(19963036);; 贵阳医学院科研基金资助(K2003-21)

作者(Author): 张婷;单可人;李毅;何燕;齐晓岚;赵艳;谢渊;吴昌学;任锡麟;官志忠;
ZHANG Ting,SHAN Ke-ren,LI Yi,HE Yan,QI Xiao-lan,ZHAO Yan,XIE Yuan,Wu Chang-xue,REN Xi-lin,GUAN Zhi-zhong(Molecular Biological Key Laboratory of Guiyang Medical College,Guiyang 550004,China)

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DOI: 10.19367/j.cnki.1000-2707.2006.01.006

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