摘要
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43~60bp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 detection of single nucleotide polymorphism(SNP) with polymerase chain reaction(PCR) in 8 patients and accuracy of the 2 sequencing techniques for the Methods: The exon 2 of Apo J gene was screened 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.
出处
《贵阳医学院学报》
CAS
2006年第1期18-20,共3页
Journal of Guiyang Medical College
基金
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)
关键词
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