NDLI: Optimization of RNA isolation and leukocyte viability in canid RNA expression studies

Content Provider	Springer Nature Link
Author	Oster, Marina Pollinger, John P. Stahler, Daniel R. Wayne, Robert K.
Copyright Year	2011
Abstract	For RNA transcript analysis, RNA source and quality influence the resolution and representation of true gene expression. We evaluate collection and purification methodologies for isolating gray wolf (Canis lupus) mRNA from peripheral whole blood to maximize detection and quantification of low expression genes. We specifically examine ambient-temperature stabilization of RNA in whole blood at time of collection, subsequent isolation and RNA stabilization of leukocytes to remove globin RNA, and removal of ribosomal RNA (rRNA) using commercially available kits. Ambient temperature RNA stabilization and globin RNA removal were successful with robust mRNA extraction yields and quality, verified by quantitative (q)PCR of reference housekeeping genes. Rhodopsin gene mRNA, present in low levels in peripheral blood, was only detected by qPCR when leukocyte mRNA was additionally treated for ribosomal RNA removal.
Starting Page	27
Ending Page	29
Page Count	3
File Format	PDF
ISSN	18777252
Journal	Conservation Genetics Resources
Volume Number	4
Issue Number	1
e-ISSN	18777260
Language	English
Publisher	Springer Netherlands
Publisher Date	2011-06-24
Publisher Place	Dordrecht
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	RNA isolation Leukoctyes Ribosomal-Depletion Expression Canids Evolutionary Biology Conservation Biology/Ecology Animal Anatomy Morphology Histology Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Genetics Ecology, Evolution, Behavior and Systematics

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