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| 基因芯片的制备研究 | |
| 引用本文： | 李瑶,陈菊祥,裘敏燕,应康,陈沁,符薇,王品,沈娴,谢毅,毛裕民.基因芯片的制备研究[J].第二军医大学学报,2000,21(9):812-814,I002. |
| 作者姓名： | 李瑶 陈菊祥 裘敏燕 应康 陈沁 符薇 王品 沈娴 谢毅 毛裕民 |
| 作者单位： | 1. 复旦大学遗传学研究所,上海,200433 2. 第二军医大学长征医院 3. 联合基因科技(集团)有限公司 |
| 基金项目： | 上海市现代生物与新药产业发展基金!资助项目( 9843 1912 1) |
| 摘 要： | 目的：建立基因芯片的制备技术。方法：基因样本准备，将带荧光标记的ＰＣＲ产物用点样仪点于玻片介质上，并经过点样后处理制备成基因芯片，用扫描仪测定前后芯片上荧光强度的变化，计算ＤＮＡ固定率，分别设计了５种玻片３种固定条件和６种点样后处理方法的固定率测定。结果：对芯片制备的各种条件和方法优化实验表明，ｃＤＮＡ的氨基修复，点样后的ＵＶ交联和芯片室温干燥处理能有效提高ＤＮＡ的固定率。结论：本研究建立的基因芯 |
| 关 键 词： | 微矩阵基因芯片 基因表达谱 DNA固定率 制备 |
Optimization and development of DNA microarray technique |
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| LI Yao,CHEN Ju-Xiang,QIU Min-Yan,YING Kang,CHEN Qin,FU Wei,WANG Pin,SHEN Xian,Xie Yi,MAO Yu-Min.Optimization and development of DNA microarray technique[J].Academic Journal of Second Military Medical University,2000,21(9):812-814,I002. | |
| Authors: | LI Yao CHEN Ju-Xiang QIU Min-Yan YING Kang CHEN Qin FU Wei WANG Pin SHEN Xian Xie Yi MAO Yu-Min |
| Abstract: | Objective:To compare and optimize conditions of DNA coupling to glass slides for development of DNA microarray. Methods: The process included preparation of DNA samples, spotting and post processing of arrays. DNA microarrays were prepared by spotting fluorescence labeled PCR products of target genes onto specially treated glass slides with robotics. The fluorescent signals before and after treatment were scanned with scanner, and the DNA attachment rate was calculated from the data obtained by ImaGene3.0 software. Results: Several key steps of this technique have been optimized, this study provided a foundation for optimizing conditions DNA attachment in the creation of DNA microarray. Conclusion: Amino modified DNA and UV cross link is useful for anchoring DNA in some distance. Drying the chip in room trmperature after spotting can enhance the DNA combination rate. |
| Keywords: | DNA microarray gene expression profile DNA attachment rate |
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