Highly efficient method for isolation of total RNA from adipose tissue.
BACKGROUND
RNA extraction is a crucial step for monitoring gene expression. Poor RNA quality (including degradation and remaining impurities) can result in misleading results. Isolation of RNA from animal tissues with high lipid content can be challenging. Especially, it is not trivial to isolate high quality RNA with a reasonable yield from adipose tissue. The aim of this study was to provide an optimized protocol for isolating total RNA from adipose tissue. This was achieved by combining the advantages of the two routinely used methods, TRI Reagent® and miRNeasy.


RESULTS
The miRNeasy method results in cleaner samples but more prone to degradation while the TRI Reagent® method results in samples contaminated with salts and solvents but more intact. The new protocol combines the best of both methods resulting in RNA of high quality and suitable for downstream experiments like RT-qPCR, microarrays and high-throughput sequencing.


CONCLUSIONS
The current protocol for total RNA isolation from adipose tissue yields sufficient amount of high quality total RNA free of contaminants.
Department of Veterinary Clinical and Animal Sciences, Section for Animal Genetics, Bioinformatics and Breeding, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg, Denmark. scs@sund.ku.dk.
Susanna
S
Cirera
D000273:Adipose Tissue / Q000737:chemistry*; D000818:Animals; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing; D020411:Oligonucleotide Array Sequence Analysis; D012313:RNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D011933:Reagent Kits, Diagnostic / Q000592:standards*; D060888:Real-Time Polymerase Chain Reaction; D013552:Swine
D011933:Reagent Kits, Diagnostic; D012313:RNA