title: Simplified and versatile method for isolation of high-quality RNA from pancreas.
abstract: Isolation of high-quality RNA from pancreas is challenging because the organ contains large quantities of RNases and undergoes autolysis upon harvest. Here we present a simplified perfusion method of the pancreas using an RNase inhibitor. The technique consistently yields high-quality RNA from stored pancreas samples suitable for molecular biology applications, including quantitative RT-PCR. Yields are comparable to RNA isolated from pancreas immediately, but superior to RNA isolated from stored samples that were snap-frozen or immersed in an RNase inhibitor solution. In addition, when compared to the previously reported in situ ductal perfusion technique, our method does not cause histological artifacts.
authors.affiliation: Department of Pediatrics, University of Iowa Carver College of Medicine, Iowa City, IA, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Griffin
authors.forename: Maisam
authors.initials: M
authors.lastname: Abu-El-Haija
authors.forename: Marwa
authors.initials: M
authors.lastname: Abu-El-Haija
authors.forename: Tatiana
authors.initials: T
authors.lastname: Rokhlina
authors.forename: Aliye
authors.initials: A
authors.lastname: Uc
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D000831:Animals, Newborn; D004791:Enzyme Inhibitors; D006652:Histological Techniques / Q000379:methods*; D051379:Mice; D010179:Pancreas / Q000737:chemistry*; D010477:Perfusion / Q000379:methods; D012313:RNA / Q000737:chemistry / Q000302:isolation & purification* / Q000592:standards; D012260:Ribonucleases / Q000037:antagonists & inhibitors; D013552:Swine
chemical_list: D004791:Enzyme Inhibitors; D012313:RNA; D012260:Ribonucleases