BioTechniques · 2012 · 33 citations · 8 references
Life SciencesBiotechnologyMolecular BiologyGene ExpressionRna BiologyTranscriptomicsRna Quality ControlRna TranscriptionAutolysis ArtifactsRibonucleic Acid ExtractionRna ExtractionRna PreservationRna Quality Assessment
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.
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Christopher S. Rogers, Yanhong Hao, Tatiana Rokhlina et al. · Journal of Clinical Investigation · 2008 · 317 citations · Full text