从Tempus管中保存的全血中同时提取mRNA和微小RNA。
Simultaneous extraction of mRNA and microRNA from whole blood stabilized in tempus tubes.
作者信息
Richards Jendai, Unger Elizabeth R, Rajeevan Mangalathu S
机构信息
Division of High-Consequence Pathogens and Pathology, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA, 30329, USA.
出版信息
BMC Res Notes. 2019 Jan 18;12(1):39. doi: 10.1186/s13104-019-4087-5.
OBJECTIVE
Studies of mRNA and miRNA expression profiling increasingly use stabilized whole blood. Commercial RNA extraction kits do not provide information about the simultaneous recovery of both mRNA and miRNA. This study evaluated yield, quality, integrity and representation of mRNA and miRNA from whole blood stabilized in Tempus tubes using three RNA extraction kits; two filter-based (Tempus and Norgen) and one bead-based (MagMax; manual vs. semi-automated, and with and without DNase treatment).
RESULTS
All RNA extraction kits and methods resulted in similar yields of mRNA (total RNA yield, quality, integrity and representation) whereas there were differences in yields of miRNA. MagMax, either manual or semi-automated, with or without DNase treatment, yielded 1.6-2.2-fold more miRNA than Tempus and Norgen kits. In addition, MagMax and Norgen methods gave greater than 12-fold more and 3.3-fold less enrichment of specific miRNA targets, respectively, in comparison to Tempus extraction reagents. This study identified MagMax kit for simultaneous recovery of both mRNA and miRNA from whole blood collected in Tempus tubes.
目的
对信使核糖核酸（mRNA）和微小核糖核酸（miRNA）表达谱的研究越来越多地使用稳定化全血。商业RNA提取试剂盒并未提供关于同时回收mRNA和miRNA的相关信息。本研究使用三种RNA提取试剂盒，评估了使用Tempus管稳定化的全血中mRNA和miRNA的产量、质量、完整性和代表性；其中两种基于过滤法（Tempus和Norgen），一种基于磁珠法（MagMax；手动与半自动，以及有无脱氧核糖核酸酶处理）。
结果
所有RNA提取试剂盒和方法产生的mRNA产量（总RNA产量、质量、完整性和代表性）相似，而miRNA产量存在差异。MagMax无论是手动还是半自动，有无脱氧核糖核酸酶处理，其产生的miRNA比Tempus和Norgen试剂盒多1.6至2.2倍。此外，与Tempus提取试剂相比，MagMax和Norgen方法分别使特定miRNA靶点的富集量增加超过12倍和减少3.3倍。本研究确定了MagMax试剂盒可用于从收集在Tempus管中的全血中同时回收mRNA和miRNA。
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