Title: Silver-staining mRNA differential display method and cloning of tumor related genes in HepG2 cell line
Abstract: AIM To develop mRNA differential display polymerize chain reaction (DD PCR) method with silver staining and to clone the liver cancer related genes in HepG2 cell line. METHODS Total RNA was extracted from the HepG2 cells and L02 cells. Their first strains of cDNA were produced by reverse transcription (RT). The cDNA were amplified by PCR using an anchored primer 5 dT11G combined with eight arbitrary primers respectively. The products of PCR were analyzed on a denaturing 60 g·L -1 polyacrylamide gel. The differential DNA bands on gel were seen by silver staining.RESULTS Several liver cancer related genes were isolated and identified by the sensitive silver staining mRNA differential display developed. CONCLUSION The established mRNA differential display method with silver staining is a simple, efficient means which has higher value for screening and cloning differentially expressed genes.
Publication Year: 2001
Publication Date: 2001-01-01
Language: en
Type: article
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