Detection of clock genes in embryoid body derived single cells with nested RT-PCR
- VernacularTitle:利用巢式聚合酶链反应检测单个胚胎小体细胞中时钟基因的表达
- Author:
Yunqian GUAN
;
Yanning CAI
- Publication Type:Journal Article
- Keywords:
embryoid body;
nested PCR;
clock genes
- From:
Basic & Clinical Medicine
2006;0(12):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish methods for examining expression of clock genes including BMAL1,CLOCK,CRY1and CRY2,and to figure out expression profile of these genes in single cell derived from embryoid body(EB).Methods Total RNA isolated from EB was subjected to reverse transcription and amplification with clock genes specific primers to determine thermo-cycle condition,which was used consequently to examine the expression profiles of these clock genes in single cells isolated with patch clamp from embryoid bodies.Results Parameters in amplification and detection were determined.No unspecific band was amplified.At least 2 molecules could be detected with established systems.In the differentiating EB cells,co-expression of these clock genes was rare.Conclusion Theses systems are sensitive enough to detect expression of clock genes in single cells.Transcription and translation loop among clock genes are not intact in differentiating EB cells,and the clock genes may play a role inthe early development and differentiation.
