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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。
Abstract：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.
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基本信息:
DOI：10.19367/j.cnki.1000-2707.2006.01.006
中图分类号:R450
引用信息:
[1]张婷,单可人,李毅,等.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[J].贵阳医学院学报,2006(01):18-20.DOI:10.19367/j.cnki.1000-2707.2006.01.006.
基金信息:
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)
2006-02-28
2006-02-28