两种氨基修饰的基因芯片表面制备方法比较

南方医科大学学报 ›› 2005, Vol. 25 ›› Issue (07): 794-798.

两种氨基修饰的基因芯片表面制备方法比较

吴清华¹, 马文丽¹, 王洪敏¹, 毛向明¹, 张宝¹, 李凌¹, 郑文岭²

1. 南方医科大学分子生物学研究所, 广东, 广州, 510515;
2. 广州军区总医院分子肿瘤学研究所, 广东, 广州, 510010

出版日期:2005-07-20 发布日期:2005-07-20
基金资助:
收稿日期:2004-12-29。
基金项目:国家自然科学基金(39880032)
作者简介:吴清华(1974- ),女,1999年硕士毕业于国防科技大学,现为第一军医大学基因工程研究所在读博士研究生,电话:020-61640114-89098,Email:wuqh@fimmu.com

Comparison of two amine-modified chemical platforms for DNA microarray preparation

WU Qing-hua¹, MA Wen-li¹, WANG Hong-min¹, MAO Xiang-ming¹, ZHANG Bao¹, LI Ling¹, ZHENG Wen-ling²

1. 南方医科大学分子生物学研究所, 广东, 广州, 510515;
2. 广州军区总医院分子肿瘤学研究所, 广东, 广州, 510010

Online:2005-07-20 Published:2005-07-20

摘要/Abstract

摘要： 目的探讨两种氨基化方法修饰玻片表面的过程及其在基因芯片制备中应用的可行性。方法玻片经清洗、硅烷化后,一种用多组分多氨化合物包被,经次亚苯基二异硫氰酸盐表面活化;另一种方法采用的包被剂是丙烯酸-丙烯酰胺单体,活化剂是碳二亚胺/N-羟基琥珀亚胺酯官能团分子。将被修饰好的玻片用于λ噬菌体基因组DNA芯片的制备,并进行杂交检测分析。结果经两种方法修饰后玻片表面分别形成分支结构和聚合物涂层,与商业化的芯片比较发现,两种芯片杂交、扫描检测后显示所得点阵的杂交信号均匀、稳定、清晰,杂交点饱满、同一性好。结论两种方法修饰的玻片用于制备基因芯片均取得了成功,其中丙烯酸-丙烯酰胺聚合物包被法处理过程简单、成本低廉、能大规模生产,是一种较理想的芯片表面制备方法。

Abstract: Objective To study two amine modification procedures for DNA microarray preparation based on polymeric coatings. Methods One of the proposed approaches utilized poly-amine coating of silanized slides activated by 1,4-phenylene diisothiocyanate,and the other employed acrylic acid-co-acrylamide copolymer and 1-(3-dimethylamino propyl)-3-ethylcar- bodiimide hydrochloride/N-hydroxysuccinimide as the coating agents and activator,respectively.The modified slides were used for preparing λ phage DNA microarrays,whose properties were analyzed by hybridization. Results Formation of dendri- meric structure and polymer was observed on the surface of the slides.The signal spots in uniform,steady and regular shape,in comparison with the commercial CMT-GAPS slides,indicated successful manufacture of the microarrays. Conclusion The two platforms are suitable for microarray preparation,and the method of acrylic acid-co-acrylamide copolymer modification is more preferred.

中图分类号:

Q789

吴清华¹, 马文丽¹, 王洪敏¹, 毛向明¹, 张宝¹, 李凌¹, 郑文岭². 两种氨基修饰的基因芯片表面制备方法比较[J]. 南方医科大学学报, 2005, 25(07): 794-798.

WU Qing-hua¹, MA Wen-li¹, WANG Hong-min¹, MAO Xiang-ming¹, ZHANG Bao¹, LI Ling¹, ZHENG Wen-ling². Comparison of two amine-modified chemical platforms for DNA microarray preparation[J]. Journal of Southern Medical University, 2005, 25(07): 794-798.

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