Highly efficient method for isolation of total RNA from adipose tissue

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• 作者：Susanna Cirera (2)
  
• 关键词：Adipose tissue ; Total RNA isolation ; RNA purity ; RNA integrity
• 刊名：BMC Research Notes
• 出版年：2013
• 出版时间：December 2013
• 年：2013
• 卷：6
• 期：1
• 全文大小：259 KB
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• 作者单位：Susanna Cirera (2)
  
  2. 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
  
• ISSN：1756-0500

文摘

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. Findings 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.