Amplification of cDNA via RT-PCR using RNA extracted from postmortem tissues.
Analysis of cDNA derived from messenger RNA is of advantage over using genomic DNA in genetic analysis of large genes, especially those with lengthy intron sequences. However, because of its instability and rapid degradation, RNA extraction from postmortem tissues has not been attempted. Here, we report the successful extraction of intact mRNA from various postmortem tissues from accidental and sudden death cases. Subsequently with reverse transcriptase-polymerase chain reaction (RT-PCR), we were able to amplify cDNA fragments of different lengths up to 0.9 kb. The described method therefore provides a useful tool in genetic analysis of postmortem tissues.
Phang
T W
TW
Department of Community, Occupational and Family Medicine, Singapore.
Shi
C Y
CY
Chia
J N
JN
Ong
C N
CN
D001483:Base Sequence; D018076:DNA, Complementary / Q000032:analysis*; D005554:Forensic Medicine / Q000379:methods*; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D009206:Myocardium / Q000737:chemistry; D016133:Polymerase Chain Reaction / Q000379:methods*; D011180:Postmortem Changes; D012333:RNA, Messenger / Q000032:analysis*; D012194:RNA-Directed DNA Polymerase
D018076:DNA, Complementary; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase