RNA interference: PCR strategies for the quantification of stable degradation-fragments derived from siRNA-targeted mRNAs.
mRNA targeted by siRNA is endogeneously cleaved into a 5'- and a 3'-fragment and finally degraded in cells. Little is known about the relative stability and degradation kinetics of these 5'- and 3'-fragments after the siRNA mediated first cut. We present a qRT-PCR protocol which allows the determination of the optimal time point for mRNA analyses, helping to avoid the generation of false positive effects in downstream experiments, such as microarray analysis, which may be caused by undegraded fragments of a siRNA-targeted mRNA.
Hahn
Peter
P
QIAGEN GmbH, QIAGEN Strasse 1, D-40724 Hilden, Germany.
Schmidt
Cornelia
C
Weber
Martin
M
Kang
Jie
J
Bielke
Wolfgang
W
D020868:Gene Silencing; D018390:Gene Targeting / Q000379:methods*; D016133:Polymerase Chain Reaction / Q000379:methods*; D034622:RNA Interference; D012333:RNA, Messenger / Q000235:genetics*; D034741:RNA, Small Interfering / Q000235:genetics*; D014162:Transfection / Q000379:methods*
D012333:RNA, Messenger; D034741:RNA, Small Interfering