Total RNA-isolation of abdominal hernia of rats for quantitative real-time reverse transcription (RT) PCR assays.
Increasing complications in incisional hernia surgery call for novel treatments. A gene expression analysis of injured tissues displays important parameters for tissue regeneration. Until today, no reliable method has been described for a quantitative gene expression analysis of hernia tissues. In this work, a protocol is described for the isolation of DNA-free total RNA of incisional hernias for the first time. Moreover, real-time RT PCR assays for collagen type I and III and TGF-beta1 are demonstrated for relative gene expression analyses. Both methods enable relative gene expression analyses of hernia tissues for the first time.
Morsczeck
Christian
C
Stiftung Caesar, Bonn, Germany. christian.morsczeck@klinik.uni-regensburg.de
Korenkov
Michael
M
Nagelschmidt
Manfred
M
Feher
Domonkos
D
Schierholz
Jörg Michael
JM
D000818:Animals; D001483:Base Sequence; D001681:Biological Assay / Q000379:methods; D024042:Collagen Type I / Q000032:analysis* / Q000235:genetics; D024061:Collagen Type III / Q000032:analysis* / Q000235:genetics; D020869:Gene Expression Profiling; D046449:Hernia, Abdominal / Q000235:genetics / Q000473:pathology* / Q000601:surgery; D012313:RNA / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D053773:Transforming Growth Factor beta1 / Q000032:analysis* / Q000235:genetics
D024042:Collagen Type I; D024061:Collagen Type III; D053773:Transforming Growth Factor beta1; D012313:RNA