Evaluation of RNA purification methods by using different blood stabilization tubes: identification of key features for epidemiological studies
Evaluation of RNA purification methods by using different blood stabilization tubes: identification of key features for epidemiological studies
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DOI：
10.1186/s13104-020-04943-4
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发表时间：
2020-02-18
影响因子：
1.8
通讯作者：
Aguilar-Quesada, R.
中科院分区：
文献类型：
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作者：
Carrillo-Avila, J. A.;de la Puente, R.;Aguilar-Quesada, R.
目的外周血是诊断和流行病学研究中最有前景的RNA生物标志物来源，因为疾病和预后信息的存在反映在基因表达模式中。高质量的RNA被许多不同的下游应用所使用，因此选择最合适的RNA稳定和纯化方法是很重要的。我们从25名献血者的300份血液样本中分析了由两名技术人员处理的RNA，使用了Tempus和Paxgene试管的三种方法。结果用TEMPUS SPIN RNA分离试剂盒和PaxGene型血液miRNA试剂盒提取的RNA质量最好，而用TEMPUS SPIN RNA分离试剂盒提取的RNA质量较高。分别用Paxgene血液miRNA试剂盒和Tempus Spin RNA分离试剂盒检测的样本中，RNA和miRNA基因的CQ值较低。我们认为TEMPUS SPIN RNA提取试剂盒是最可靠的方法，而MAGMAX稳定血管RNA提取试剂盒显示最不稳定。对于处理大量队列并进行流行病学研究的生物库来说，Tempus Spin RNA分离试剂盒是最合适的方法。这项研究证明了现实生活中程序的健壮性。
Objective Peripheral blood is the most promising source of RNA biomarkers for diagnostic and epidemiological studies, because the presence of disease and prognostic information is reflected in the gene expression pattern. Quality RNA is used by a number of different downstream applications, so the selection of the most appropriate RNA stabilization and purification method is important. We have analyzed the RNA purified from 300 blood samples from 25 donors processed by two technicians using three methodologies with Tempus and PaxGene tubes. Results The best quality sample results were obtained with the Tempus Spin RNA Isolation Kit and the PaxGene Blood miRNA Kit, although larger amounts of RNA were obtained with the Tempus Spin RNA Isolation Kit. Lower Cq values were observed for RNA and miRNA genes in samples that were tested with PaxGene Blood miRNA Kit and Tempus Spin RNA Isolation Kit respectively. We identify the Tempus Spin RNA Isolation Kit as the most robust methodology, whilst the MagMax for Stabilized Blood Tubes RNA Isolation Kit showed the most instability. For biobanks, which process a large cohort and conduct epidemiological studies, the Tempus Spin RNA Isolation Kit is the most appropriate methodology. The study demonstrates the robustness of real-life procedures.