基于寡核苷酸芯片的引物连接法的研究  

Research of Ligase Catalyzed Primer Ligation Based on the Oligonucleotide Microarray

作　　者：章承芝[1] 张红发 李瑶[1] 曹慧敏 韩志勇 裘敏燕 谢毅[1] 毛裕民[1] 

机构地区：[1]复旦大学生命科学学院遗传学研究所,上海200433 [2]上海博星基因芯片有限公司,上海200092

出　　处：《复旦学报（自然科学版）》2003年第4期608-613,620,共7页Journal of Fudan University：Natural Science

摘　　要：为了更好地将寡核苷酸芯片应用于单个核苷酸差异的检测,以HBV质粒为模板,探索了基于寡核苷酸芯片的引物延伸法.对主要反应体系和条件进行了优化:将杂交和连接的过程合并,创立了一步法,反应条件为30℃1h;在体系中加入了TritonX 100(φ=0.0067%),大大降低了背景信号.另外,将引物连接一步法应用于ABO血型分型,证明了其准确、有效,能够应用于分析单个核苷酸的差异情况.Ligase catalyzed primer ligations based on the oligonucleotide microarray was investigated in order to better detect single nuleotide diversity. The HBV fragment from the plasmid was used as a template. Ligase catalyzed ligations were optimized in some main conditions: adding 0.006 7% Triton X100, so as to reduce the backgound noise; combining the two step of hybridization and ligation and initiating the first onestep method. In addition, the optimized protocal was applied in detecting the blood genotype ABO, which proved that this method was accurate and efficient to analyze the specific nucleotide diversity and can also be used in the disease diagnosis, genotyping and mutation detection.

关 键 词：寡核苷酸芯片 引物连接法 引物连接一步法 

分 类 号：Q75[生物学—分子生物学]