[The degradation of housekeeping mRNA in dead rats by real-time RT-PCR].
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.


CONCLUSIONS
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.
Ren
Guang-mu
GM
School of Forensic Medicine, Shanxi Medical University, Taiyuan 030001, China. renguangmu@163.com
Liu
Ji
J
Wang
Ying-yuan
YY
D000199:Actins / Q000235:genetics*; D000818:Animals; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000235:genetics*; D011180:Postmortem Changes; D020871:RNA Stability / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013997:Time Factors
D000199:Actins; D012333:RNA, Messenger; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases