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消减杂交技术结合限制性显示技术制备K562细胞特异基因探针

Preparation of Specific Gene Probes of K562 Cells with Subtraction Hybridization and Restriction Display

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【作者】 宋艳斌; 马文丽; 冯春琼; 石嵘; 毛向明; 张宝; 郑文岭;

【Author】 SONG Yan-bin1,MA Wen-li1,FENG Chun-qiong1,SHI Rong1, MAO Xiang-ming1,ZHANG Bao1,ZHENG Wen-ling2(1.Institute of Molecular Biology,Nanfang Medical University,Guangzhou 510515,Guangdong,China; 2.Huanan Biochip Center,Guangzhou 510515,Guangdong,China)

【机构】 南方医科大学分子生物学研究所; 华南生物芯片中心 中国广东广州510515; 中国广东广州510515; 中国广东广州510010;

【摘要】 建立一种简便、快速、特异的制备基因芯片探针的方法.以K 562细胞和正常人淋巴细胞作为消减对象,利用自行建立的消减方法进行消减杂交,结合限制性显示技术,分组扩增差异cD N A,回收K 562细胞特异基因片段,制作基因芯片探针.结果显示,分离到400个K 562特异的基因,片段大小均一,适于制作cD N A芯片.消减杂交技术结合限制性显示技术制备基因芯片探针,具有快速、简便、特异的特点,降低了芯片制作成本,可加速芯片的推广应用.

【Abstract】 A fast,easy and specific probe preparation method of microarray was s tudied.Double-strand cDNA from K562 cells(tester) and normal lymphocyte(driv er) were synthesized by reverse transcription respectively.After three times of subtractive hybridization and polymerase chain reaction(PCR) amplification in subgroups with restriction display,the cDNA fragments were isolated by PAGE and secondary amplified to prepare probes of microarray.400 specific cDNA fragments of K562 cells were obtained as probes of microarray using subtraction hybridiza tion and restriction display.The probes length is comparatively homogeneous and suitable for microarray fabrication.Combination of subtraction hybridization an d restriction display technology could provide a fast,easy and specific method f or probe-preparation of microarray.

【基金】 国家自然科学基金资助项目(39880032)
  • 【文献出处】 生命科学研究 ,Life Science Research , 编辑部邮箱 ,2005年03期
  • 【分类号】R346;
  • 【被引频次】1
  • 【下载频次】59
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