Sarkar F H, Li Y W, Ball D E, Crissman J D
Department of Pathology, Wayne State University, School of Medicine, Detroit, MI.
Biotechniques. 1992 Jan;12(1):22, 24, 26.
During the development of a micro-method for the quantitative analysis of gene expression, we observed that the sensitivity for detection of PCR-amplified product was higher when silver staining was used, compared with either ethidium bromide staining (as reported previously) or Southern hybridization with 32P-labeled oligonucleotide probe. This observation may have an important impact on the analysis of RNA-PCR-amplified signal for the quantitation of mRNA expression, simply by densitometric scanning of silver-stained polyacrylamide gel. We believe that this is the first report to show such a comparison, demonstrating the greater sensitivity of silver staining compared with either ethidium bromide staining or Southern hybridization utilizing an oligonucleotide probe prepared by 3'-end labeling with 32P-dATP.
在开发一种用于基因表达定量分析的微量方法过程中,我们观察到,与溴化乙锭染色(如先前报道)或用32P标记的寡核苷酸探针进行Southern杂交相比,使用银染色时,对PCR扩增产物的检测灵敏度更高。这一观察结果可能对通过银染聚丙烯酰胺凝胶的光密度扫描来定量mRNA表达的RNA-PCR扩增信号分析产生重要影响。我们认为这是第一份展示此类比较的报告,证明了银染色比溴化乙锭染色或使用通过3'-末端用32P-dATP标记制备的寡核苷酸探针进行的Southern杂交具有更高的灵敏度。