Highly efficient method for isolation of total RNA from adipose tissue.
Highly efficient method for isolation of total RNA from adipose tissue.
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DOI：
10.1186/1756-0500-6-472
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发表时间：
2013-11-18
影响因子：
1.8
通讯作者：
Cirera S
中科院分区：
文献类型：
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作者：
Cirera S
RNA提取是监测基因表达的关键步骤。较差的RNA质量（包括降解和残留的杂质）可能导致误导性的结果。从脂质含量高的动物组织中分离RNA是具有挑战性的。特别是，从脂肪组织中分离出高质量的RNA并不是一件容易的事。本研究的目的是提供一种从脂肪组织中分离总RNA的优化方案。这是通过结合TRI Reagent®和miRNeasy两种常规方法的优势来实现的。miRNeasy方法得到的样品更干净，但更容易降解，而TRI试剂®方法得到的样品被盐和溶剂污染，但更完整。新方案结合了两种方法的优点，产生高质量的RNA，适用于RT-qPCR，微阵列和高通量测序等下游实验。目前从脂肪组织中分离总RNA的方案产生足够数量的无污染物的高质量总RNA。
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. 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. The current protocol for total RNA isolation from adipose tissue yields sufficient amount of high quality total RNA free of contaminants.