Library construction of HepG2 cells by serial analysis of gene expression
10.3760/cma.j.issn.1673-9752.2009.02.016
- VernacularTitle:利用基因表达连续分析技术构建肝癌HepG2细胞文库
- Author:
Weihui LIU
;
Ren LI
;
Kefeng DOU
- Publication Type:Journal Article
- Keywords:
Carcinoma,hepatocellular;
HepG2 cells;
Serial analysis of gene expression;
Library
- From:
Chinese Journal of Digestive Surgery
2009;8(2):120-123
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a sequence tag library of HepG2 cells by serial analysis of gene expression (SAGE). Methods Total RNA of HepG2 cells was extracted using a Trizol agent. The double strand cDNA were reverse-translated from mRNA which was isolated from total RNA. According to the protocol of SAGE, a sequence tag library of HepG2 cells was constructed. The transcript, which was represented by each tag, was analyzed by consulting the GenBank and UniGene, and the abundance of the tags was analyzed by SAGE analysis software. Results A total of 14 181 SAGE tags out of 15 586 tags were steadily and effectively amplified and the lengths of the tags were between 200-3000 bp. After sequence analysis of the tags, 2023 specific genes were revealed. Conclusions The use of improved reverse transcription agent kit may guarantee obtaining the full length cDNA, which provides materials for the construction of SAGE library. SAGE is a powerful method to investigate the gene expression profile, and it establishes a foundation for gene researches.
