The degradation of housekeeping mRNA in dead rats by real-time RT-PCR.
- Author:
Guang-mu REN
1
;
Ji LIU
;
Ying-yuan WANG
Author Information
1. School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China. renguangmu@163.com
- Publication Type:Research Support, Non-U.S. Gov't
- MeSH:
Actins/genetics*;
Animals;
Glyceraldehyde-3-Phosphate Dehydrogenases/genetics*;
Postmortem Changes;
RNA Stability/genetics*;
RNA, Messenger/metabolism*;
Rats;
Rats, Sprague-Dawley;
Reverse Transcriptase Polymerase Chain Reaction/methods*;
Time Factors
- From:
Journal of Forensic Medicine
2009;25(1):33-36
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To explore the feasibility of real-time RT-PCR of housekeeping mRNA degradation in dead rats in order to seek new suitable techniques for post-mortem interval (PMI).
METHODS:The levels of GAPDH mRNA and beta-actin mRNA in the brain and spleen of the rats were measured at different times after death by the SYBR Green I real-time RT-PCR. The relative mRNA level was indicated with the Ct value, then the relationship between the Ct value and PMI were analyzed and the corresponding regression equation was obtained.
RESULTS:The Ct values of GAPDH mRNA and beta-actin mRNA by SYBR Green I real time RT-PCR correlated well with the PMI.
CONCLUSION:The SYBR Green I real-time RT-PCR is a reliable technique for studying mRNA degradation. Adoption of the housekeeping genes eliminates systematical errors of other genes which have individual difference. As an objective and dynamic indication, Ct value has a good correlation with PMI, and could be applied for estimating PMI, especially in late period.
