Real-time quantitative RT-PCR for the detection of bcl-2 mRNA expression.
- Author:
Zhaoxiu ZHANG
1
;
Lianhuang LU
;
Xiaopeng LAN
;
Jianda HU
;
Minhui LIN
;
Quansheng JING
Author Information
- Publication Type:Journal Article
- MeSH: Electrophoresis, Agar Gel; methods; Gene Expression; Humans; Proto-Oncogene Proteins c-bcl-2; genetics; RNA, Messenger; analysis; Reverse Transcriptase Polymerase Chain Reaction
- From: Chinese Journal of Medical Genetics 2002;19(5):412-415
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo establish a real-time quantitative RT-PCR method for the detection of expression of bcl-2 mRNA.
METHODSThe vector containing bcl-2 gene as standard template was constructed with T-A cloning technique. The fluorogenic probe (i.e.,Taq man probe) was used to establish a real-time RT-PCR. bcl-2 mRNA expression level in Burkitt's lymphoma pre-and post-treated with bcl-2 antisense phosphothioate oligodeoxynucleotides (AS-PS-ODN) was determined with real-time quantitative RT-PCR, also with semi-quantitative RT-PCR.
RESULTSThe expression of bcl-2 mRNA in Burkitt's lymphoma treated with bcl-2 AS- PS-ODN decreased significantly and no changes of bcl-2-mRNA expression in group treated with nonsense ODN were noticed. The semi-quantitative RT-PCR results also demonstrated that bcl-2 level varied as detected with real-time fluorogenic quantitative RT-PCR, but less sensitive and accurate.
CONCLUSIONDetection of bcl-2 mRNA expression with the fluorogenic real-time quantitative RT-PCR is more accurate and sensitive than semi-quantitive RT-PCR.
