摘要
应用ＲＡＰＤ－ＰＣＲ技术结合地高辛非放射性标记系统制备了人巨细胞病毒ＤＮＡ探针，并与常规随机引物法进行对照。ＲＡＰＤ－ＰＣＲ制备的ＨＣＭＶＤＮＡ探针长度呈多态性，扩增过程中不需合成特异引物，探针特异性强，产量高，用５０ｎｇＨＣＭＶＤＮＡ模板即可制备８．０μｇ探针；标记体系中ＲＡＰＤ－ＰＣＲ方法Ｄｉｇ－ｄＵＴＰ浓度为５．２％。表明ＲＡＰＤ－ΡＣＲ技术可用于少量ＤＮＡ模板制备大量ＤＮＡ探针，适用于原位杂交等需高浓度探针的核酸杂交及临床大量标本的测试。
The application of RAPD PCR in labeling HCMV DNA probe combined with digoxingen labeling and detection kit was performed.The yield of HCMV DNA probe labeled with RAPD PCR was higher than that labeled with routinal random primer labeling method,8000ng vs 800ng detected by direct dot blot.The sensitivity of HCMV DNA probe labeled with RAPD PCR was higher than that labeled with routinal method,0.1 pg vs 0.3 pg,detected by homologous dot blot hybridization.The DNA probe labeled with RAPD PCR was polymorphic.These DNA probes suited to different nucleic acid hybridization,especially in situ hybridization.
出处
《山东医科大学学报》
1998年第2期118-120,共3页
Acta Academiae Medicinae Shandong
基金
国家自然科学基金
关键词
巨细胞病毒
地高辛
随机扩增
多态DNA
Cytomegaloviruses
Probe
DNA
Digoxingen
Randomly amplified polymorphic DNA
