在载玻片上制备高杂交效率的DNA芯片  

Preparation of high-efficiently hybridization DNA chip on glass support

作　　者：蒋笃孝[1] 宋龄瑛[2] 王德育[3] 袁春伟[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3期65-70,共6页Journal of Jinan University(Natural Science & Medicine Edition)

基　　金：国务院侨办重点科研项目基金! (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资助!(2 0 0 0B7)

摘　　要：目的 :改进玻璃基质上固定DNA的方法 ,提高固定DNA的杂交效率 .方法 :玻片用 β -(对、间氯甲基苯基 )三氯硅乙烷和 β -苯基三氯硅乙烷二组分混合硅烷化试剂处理 ,再用NaI丙酮溶液处理 ,在表面自组装形成带有苄碘功能基的二组分混合单层硅烷膜 .巯基修饰的DNA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的优化密度为9 3× 10 - 2 μmol/m2 ,目标DNA杂交密度为 8.6× 10 - 2 μmol/m2 ,固定DNA的杂交率为 93% .结论 :采用两步竞争性反应的方法能够较容易地控制DNA在芯片上的固定密度 ,固定的DNA在芯片分布较均匀 ,因此制得的DNA芯片具有较高杂交效率和杂交密度 。Aim: A method for the covalent binding of oligonucleotides to solid glass supports was developed . Methods: Glass wafers were derivatized with β-phenethyltrichlorosilane and 1-trichlorosily-2-(p, m-chloromethylphenyl)- ethane, followed by treatment with acetone solution of NaI to create self-assembled mixed monolayer silane films with iodomethylphenyl group. Thio-derivatized, single-stranded DNA was then coupled to the iodomethylphenyl surface in simple one-step reaction. Results: The optimal surface density was 9.3×10 -2 μmol of DNA/m 2. 93% of the probes were available for hybridization. Conclusion: The two steps of competitive reaction may ensure that the probes are nearly uniformly distributed on wafer surface. This system has highly efficient hybridization, and can be conveniently applied for studies on hybridization and detection of nucleic acids.[

关 键 词：DNA 共价键固定 玻璃基质 高杂交效率 核酸杂交 杂交密度 硅烷膜 

分 类 号：Q781[生物学—分子生物学]