摘要
目的:检测白种人和汉族人RDH8基因序列中单核苷酸多态性(SNP)位点,探讨变性高效液相色谱分析法(DHPLC)在基因组研究中的效能。方法:建立8个由白种人和汉族人基因组DNA组成的混合样品池,并进行PCR扩增。其产物以DHPLC检测,发现杂合子信号后,对单独DNA样本进行DHPLC检测及直接测序确定其突变类型。以DHPLC结合DNA样本混合技术在150个汉族人和20个欧洲白种人样本中测定SNP的基因频率。结果:在RDH8基因中共检测到17个SNP,其中12个为新发现的,5个为已报道的;11个SNP在筛查阶段发现,6个在基因频率检测阶段发现;2个仅在白种人样本中检测到,3个SNP的MAF在两种人种间有较明显的差异;汉族人基因频率检测确定了7个常见SNP。结论:DHPLC能有效地用于基因组SNP的检测,结合DNA混合技术,更能提高其检测效力,有效地应用于基因频率的测定。
<Abstrcat> Objective: To assess the efficacy of denaturing high-performance liquid chromatography (DHPLC) as a new screening assayfor sequence variation in SNP detecting and new approach to SNP genotyping. Methods: DHPLC was used to screen SNPs in 8 DNA pools each consisting of DNA from 5 individuals,and genotype the identified SNPs in 150 Chinese subjects from Hong Kong. Results: Seventeen SNPs were identified:12 were novel and 5 were previously reported; 11 were found in screening stage and the other 6 were found in genotyping stage; 2 were only found in Caucasian samples; 3 showed ethnic difference of minor allele frequency(MAF); 7 were common with the MAF>0.05 in Chinese samples. Conclusion: The results demonstrated the efficiency of DHPLC in screening SNPs when coupled with DNA pooling strategy,and in genotyping SNPs.
出处
《浙江大学学报（医学版）》
CAS
CSCD
2005年第3期248-254,共7页
Journal of Zhejiang University(Medical Sciences)
