Ferraro Davide, Champ Jérôme, Teste Bruno, Serra M, Malaquin Laurent, Descroix Stéphanie, de Cremoux Patricia, Viovy Jean-Louis
Macromolecules and Microsystems in Biology and Medicine, Institut Curie, Centre National de la Recherche Scientifique, Université Pierre et Marie Curie, PSL Research University, UMR 168, 75005, Paris, France.
University Paris-Diderot, PRES Paris Cité, INSERM/CNRS UMR944/7212, Paris, France.
Methods Mol Biol. 2017;1547:113-121. doi: 10.1007/978-1-4939-6734-6_9.
Analyses of nucleic acids are routinely performed in hospital laboratories to detect gene alterations for cancer diagnosis and treatment decision. Among the different possible investigations, mRNA analysis provides information on abnormal levels of genes expression. Standard laboratory methods are still not adapted to the isolation and quantitation of low mRNA amounts and new techniques needs to be developed in particular for rare subsets analysis. By reducing the volume involved, time process, and the contamination risks, droplet microfluidics provide numerous advantages to perform analysis down to the single cell level.We report on a droplet microfluidic platform based on the manipulation of magnetic particles that allows the clinical analysis of tumor tissues. In particular, it allows the extraction of mRNA from the total-RNA sample, Reverse Transcription, and cDNA amplification, all in droplets.
医院实验室经常进行核酸分析，以检测基因改变，用于癌症诊断和治疗决策。在各种可能的检测中，mRNA分析可提供有关基因表达异常水平的信息。标准实验室方法仍不适用于低mRNA量的分离和定量，特别是对于罕见亚群分析，需要开发新技术。通过减少涉及的体积、处理时间和污染风险，微滴微流控技术为进行单细胞水平的分析提供了许多优势。我们报道了一种基于磁性颗粒操纵的微滴微流控平台，该平台可用于肿瘤组织的临床分析。特别是，它允许在液滴中从总RNA样本中提取mRNA、进行逆转录和cDNA扩增。
