mRNA差异显示技术中差异条带的回收与再扩增  

Recovering and Reamplifying the Differentially Expressed cDNA Bands Isolated from mRNA Differential Display

作　　者：王宏[1,2,3,4] 李春海[1,2,3,4] 陈高明[1,2,3,4] 王尧河 夏贞彪[1,2,3,4] 杨毅 

机构地区：[1]军事医学科学院附属医院肿瘤分子生物学实验室 [2]河南医科大学病理学教研室 [3]军事医学科学院放射医学研究所 [4]天津医科大学附属肿瘤医院分子生物学实验室

出　　处：《中国肿瘤临床》1999年第5期350-351,共2页Chinese Journal of Clinical Oncology

基　　金：国家自然科学基金

摘　　要：目的：对来源于两种或两种以上组织的ＲＮＡ经反转录和ＰＣＲ后进行比较。方法：采用ｍＲＮＡ差异显示过程中差异条带的回收与再扩增的方法。结果：直接将切下的凝胶条简单洗涤和碾碎后制备的再扩增模板比传统的糖原沉淀法简单有效。结论：再扩增过程中将３０循环的低退火温度的ＤＤＰＣＲ和２０循环的严格条件的ＰＣＲ结合，能获得较高的再扩增效率。Methods for retrieving and reamplifying the differentially expressed cDNA bands have been modified Direct reamplification of differentially expressed bands after cutting from a PAG followed by a simple rinse and crush step has proved to be more convenient and effective than the traditional glycogen-precipitation method Combination of 30 cycles of differential display polymerase chain reaction (DDPCR) and 20 cycles of standard PCR reaction also yielded higher reamplification rates

关 键 词：MRNA差异显示 差异条带回收 再扩增 肿瘤 

分 类 号：R730.43[医药卫生—肿瘤]