一种玻璃载片共价键固定DNA的新方法  

A novel method for covalent attachment of synthetic DNA to glass support

作　　者：蒋笃孝[1] 宋岭瑛[2] 王德育[3] 

机构地区：[1]暨南大学化学系,广东广州510632 [2]广州师范学院化学系,广东广州510400 [3]东南大学分子与生物分子电子学国家重点研究实验室,江苏南京210018

出　　处：《暨南大学学报（自然科学与医学版）》2001年第1期78-83,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国务院侨办重点科研基金!项目 (93A119) ;东南大学分子与生物分子电子学国家重点实验室开放课题基金!(2 0 0 0B7)资助

摘　　要：目的 :建立了一种简单、快速、高效共价固定DNA到玻璃基质上的方法 .方法 :玻片用β - (对、间氯甲基苯基 )三氯硅乙烷处理在表面自组装形成带有苄氯功能基的单层硅烷膜 ,然后再用NaI丙酮溶液处理将苄氯基转化为苄碘基 .巯基修饰的DNA通过巯基与苄碘基反应形成硫醚键共价偶联到这一自组装的硅烷膜上 结果 :放射性分析证实玻璃基质表面共价固定DNA的密度为3.2nmol/m2 .用放射性标记的目标DNA进行杂交试验表明最大杂交密度为 0 77nmol/m2 .结论 :本方法只需一步偶联反应 ,反应时间短、偶联效率高、操作简单、不需要严格的试剂 ,而且制得的DNA芯片有较高的温度稳定性 ,可方便地用于核酸杂交和探测研究 .Aim: A simple, rapid, and efficient method for the covalent binding of thio-modified DNA oligomers to glass supports was developed. Methods: Glass slides were derivatized with 1-trichlorosily-2-(p, m-chloromethylphenyl)ethane, and then treated with acetone solution of NaI to create self-assembled monolayer silane films with iodomethylphenyl group. An oligonucloetide containing a 3'- thiol was then coupled to the -CH 2 I surface by reacting the thiol groups with -CH 2I. Results:Analysis of the surface reveals that the mount of covalently bound oligonucloetides is 3.2 nmol of DNA/m 2. Hybridization with radioactively labeled oligonuloetide probes showed that the density of hybridization was 0 77 nmol of DNA/m 2.Conclusion: The method reported here has the advantage that HS-modified DNA is bound to the iodomethylphenyl fuctionality-derivatized glass directly in simple one step. Time-consuming reactions, difficult manipulations, and critical reagents are avoided. Moreover, the covalently bound DNA described here have shown high thermal stability. This system can be conveniently applied for studies on hybridization and detection of nucleic acids.[

关 键 词：脱氧寡核苷酸 共价键固定 玻璃基质 DNA 硅烷膜 偶联反应 生物传感器 

分 类 号：Q782[生物学—分子生物学]