Application of in situ ductal perfusion to facilitate isolation of high-quality RNA from mouse pancreas.
A technique to isolate high-quality intact RNA from murine pancreas is described. This technique involves in situ ductal perfusion of the pancreas with an RNase inhibitor prior to removal of the organ for RNA extraction. In this way, the pancreatic RNases are inhibited in situ allowing good yields of intact RNA, suitable for studies on pancreatic gene transcription by real-time PCR or microarray analysis, to be obtained in a reliable way.
Mullin
Anne E
AE
University of British Columbia, Vancouver and Child and Family Research Institute, Vancouver, BC, Canada.
Soukatcheva
Galina
G
Verchere
C Bruce
CB
Chantler
Janet K
JK
D000818:Animals; D051379:Mice; D008815:Mice, Inbred Strains; D008953:Models, Anatomic; D010179:Pancreas / Q000737:chemistry* / Q000601:surgery; D010183:Pancreatic Ducts / Q000502:physiology*; D010477:Perfusion / Q000379:methods*; D012313:RNA / Q000302:isolation & purification*; D012260:Ribonucleases / Q000037:antagonists & inhibitors*
D012313:RNA; D012260:Ribonucleases