摘要
目的:比较采用常规PCR和不对称PCR分别进行样品的掺入荧光标记,应用于60mer寡核苷酸芯片杂交,分析其对芯片杂交效率的影响。方法:以A/T克隆分别将NS1、NS2a、NS2b、NS4a和NS4b基因片段与pMD18-T载体连接,应用常规PCR和不对称PCR两种方法进行掺入荧光标记,将荧光标记样品与寡核苷酸芯片杂交,在相同条件下完成杂交清洗和芯片的扫描检测。结果:与常规PCR标记方法相比,采用不对称PCR技术标记单链样品与寡核苷酸芯片杂交,能提高芯片的杂交效率。结论:应用不对称PCR技术对样品进行荧光标记后,与寡核苷酸芯片杂交能提高芯片的杂交效率,有利于检测型寡核苷酸基因芯片的应用。
Objective：To study the hybridization efficiency of two fluorescence labeling methods by oligo microarray hybridization. nethod：Gene fragments of NS1,NS2a, NS2,NS4a and NS4b were ligated with pMD18-T vector by A/T clone. Then they were labeled by traditional PCR and asymmetric PCR. The labeled samples were examined by the oligo microarray, followed by washing and scanning under the same conditions. Results： Comparing to the traditional PCR labeling method, the asymmetric PCR labeling method showed superior results with enhanced hybridization efficiency. Conclusion：The hybridization efficiency can be enhanced by asymmetric PCR labeling method, which improving the applicability of oligo microarray technology.
出处
《军医进修学院学报》
CAS
北大核心
2005年第4期266-268,共3页
Academic Journal of Pla Postgraduate Medical School
基金
国家自然科学基金资助项目(39880032)
作者简介
张海燕（1981-），女，安徽来安人，南方医科大学在读硕士研究生（原第一军医大学）。电话：（020）61648114-89097
