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Simplified and Versatile Method for Isolation of High-Quality RNA from Pancreas
Author(s) -
Michelle Griffin,
Maisam AbuElHaija,
Marwa Abu El Haija,
Tatiana Rokhlina,
Aliye Uç
Publication year - 2012
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/0000113862
Subject(s) - rna , rnase p , pancreas , biology , rna extraction , isolation (microbiology) , microbiology and biotechnology , biochemistry , chemistry , gene , bioinformatics
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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