Simplified and versatile method for isolation of high-quality RNA from pancreas

Biotechniques. 2012 May;52(5):332-4. doi: 10.2144/0000113862.

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.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Analysis of Variance
  • Animals
  • Animals, Newborn
  • Enzyme Inhibitors
  • Histological Techniques / methods*
  • Mice
  • Pancreas / chemistry*
  • Perfusion / methods
  • RNA / chemistry
  • RNA / isolation & purification*
  • RNA / standards
  • Ribonucleases / antagonists & inhibitors
  • Swine

Substances

  • Enzyme Inhibitors
  • RNA
  • Ribonucleases