摘要
原位聚合酶链反应（ｉｎｓｉｔｕＰＣＲ）是一项高新生物技术，它不仅可以检测在细胞中微量的ＤＮＡ或者ＲＮＡ，而且可以精确定位，因此在分子病理学、分子生物学、分子遗传学和微生物学等领域都有重要的应用价值。本文根据最新的研究资料，介绍了这项技术的基本原理、分类、主要实验步骤和注意事项，并建议在间接原位聚合酶链反应（ｉｎｄｉｒｅｃｔｉｎｓｉｔｕＰＣＲ）中用引物原位标记（ＰＲＩＮＳ）技术代替荧光标记原位杂交（ＦＩＳＨ）检测信号。
Since 1990,researchers have proposed performing polymerase chain reactions(PCR)in situ in nondisrupted cells,tissue and embryo sections with subsequent fluorescence in situ hybridization(FISH)detection of the amplicon at the site of origin.The methodology termed in situ PCR has been widely applied to detect viral DNA,single copy genes and gene rearrangements in cell suspensions and fixed tissue preparations,and to localise low copy sequences in inter/metaphase chromosomes in the fields of molecular biology,genetics,pathology,medicine and microbiology.The overview described development,classification,general protocols and applications of in situ PCR technique.Using primed in situ labelling (PRINS)instead of FISH to detect the signals in the indirect in situ PCR was recommended. The common problems and relevant remedies were discussed,which may provide an useful guide for adoption of this technique.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
1999年第2期94-96,共3页
Journal of Nanjing Agricultural University
关键词
聚合酶链反应
原位聚合酶
荧光标记
原位杂交
polymerase chain reaction(PCR)
in situ PCR
fluorescence in situ hybridization(FISH)
