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从单个PAXgene血液RNA管中分离微阵列级总RNA、微小RNA和DNA。

Isolation of microarray-grade total RNA, microRNA, and DNA from a single PAXgene blood RNA tube.

作者信息

Kruhøffer Mogens, Dyrskjøt Lars, Voss Thorsten, Lindberg Raija L P, Wyrich Ralf, Thykjaer Thomas, Orntoft Torben F

机构信息

Brendstrupgaardsvej 102, DK-8200 Aarhus N, Denmark.

出版信息

J Mol Diagn. 2007 Sep;9(4):452-8. doi: 10.2353/jmoldx.2007.060175. Epub 2007 Aug 9.

DOI:10.2353/jmoldx.2007.060175
PMID:17690207
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1975097/
Abstract

We have developed a procedure for isolation of microRNA and genomic DNA in addition to total RNA from whole blood stabilized in PAXgene Blood RNA tubes. The procedure is based on automatic extraction on a BioRobot MDx and includes isolation of DNA from a fraction of the stabilized blood and recovery of small RNA species that are otherwise lost. The procedure presented here is suitable for large-scale experiments and is amenable to further automation. Procured total RNA and DNA was tested using Affymetrix Expression and single-nucleotide polymorphism GeneChips, respectively, and isolated microRNA was tested using spotted locked nucleic acid-based microarrays. We conclude that the yield and quality of total RNA, microRNA, and DNA from a single PAXgene blood RNA tube is sufficient for downstream microarray analysis.

摘要

我们开发了一种从保存在PAXgene Blood RNA管中的全血中分离微小RNA、基因组DNA以及总RNA的方法。该方法基于在BioRobot MDx上的自动提取，包括从一部分稳定化血液中分离DNA以及回收否则会丢失的小RNA种类。这里介绍的方法适用于大规模实验，并且易于进一步自动化。所获得的总RNA和DNA分别使用Affymetrix Expression和单核苷酸多态性基因芯片进行检测，分离出的微小RNA使用基于锁核酸的点阵微阵列进行检测。我们得出结论，来自单个PAXgene血液RNA管的总RNA、微小RNA和DNA的产量和质量足以用于下游微阵列分析。

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