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结核杆菌检测基因芯片的制备研究

Research on Preparation of Mycobacterium Tuberculosis DNA Microarray

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【作者】 张万江鲍朗王晓樱谢勇恩陈玮张会东于向华

【Author】 Zhang Wanjiang, Bao Lang, Wang Xiaoying, Xie Yong’en, Chen Wei, Zhang Huidong, Yu Xianghua. Research Unit of Infection and Immunity, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China

【机构】 四川大学华西基础医学与法医学院感染免疫研究室四川大学华西基础医学与法医学院

【摘要】 目的 建立结核杆菌检测基因芯片的制备技术。方法 制备和标记靶基因 ,用点样仪将靶基因点于玻片介质上 ,并经点样后处理制成基因芯片 ,用扫描仪测定处理前后芯片上荧光强度的变化 ,计算 DNA固定率 ,同时测定和比较两种不同玻片、不同点样液和三种不同点样后处理方法的 DNA固定率。结果 制备结核杆菌检测基因芯片 ,用醛基修饰玻片作为介质 ,用 DMSO溶液作为点样液 ,点样后芯片处理以水合 1小时再干燥 30分钟为佳。结论 本研究所建立的结核杆菌检测基因芯片制备技术具有较高的效率和可靠的实用性能

【Abstract】 Objective To optimize and develop the technique for mycobacterium tuberculosis DNA microarray.Methods The process included preparation of DNA samples, spotting and past-spotting treatment 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 a scanner, and the DNA attachment rate was calculated from the obtained data by software.Results A foundation for optimizing the conditions of Mycobacterium tuberculosis DNA microarrays has been laid . The support aldehyde-modified glass slide is useful for anchoring DNA at Some distance. DMSO as spotting solution is of benefit to preparation of Mycobacterium tuberculosis DNA microarray. Drying the chip at 37℃ temperature after spotting can enhance the DNA combination rate. Conclusion Several key steps of this technique have been optimized. This study has provided a foundation for optimizing the DNA attachment conditions in creating mycobacterium tuberculosis DNA microarray.

【基金】 四川省学术和技术带头人培养基金资助项目 (项目编号498992 3)
  • 【文献出处】 华西医科大学学报 ,Journal of West China University of Medical Sciences , 编辑部邮箱 ,2002年02期
  • 【分类号】R346
  • 【被引频次】6
  • 【下载频次】192
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