使用被动液滴融合技术对KRAS突变进行多重检测。

Multiplex Detection of KRAS Mutations Using Passive Droplet Fusion.

作者信息

Pekin Deniz, Taly Valerie

机构信息

CNRS, Univ. Bordeaux, CRPP, UPR 8641, 115 Avenue Schweitzer, F-33600, Pessac, France.

INSERM UMRS1147, CNRS SNC 5014, Université Paris Descartes, Equipe labellisée Ligue Nationale contre le cancer 2016, 45 rue des Saints-Pères, 75270, Paris Cedex 06, France.

出版信息

Methods Mol Biol. 2017;1547:133-142. doi: 10.1007/978-1-4939-6734-6_11.

DOI:10.1007/978-1-4939-6734-6_11

PMID:28044293

Abstract

We describe a droplet microfluidics method to screen for multiple mutations of a same oncogene in a single experiment using passive droplet fusion. Genomic DNA from H1573 cell-line was screened for the presence of the six common mutations of the KRAS oncogene as well as wild-type sequences with a detection efficiency of 98 %. Furthermore, the mutant allelic fraction of the cell-line was also assessed correctly showing that the technique is quantitative.

摘要

我们描述了一种液滴微流控方法，可在单个实验中使用被动液滴融合来筛选同一癌基因的多个突变。对H1573细胞系的基因组DNA进行了KRAS癌基因六个常见突变以及野生型序列的筛选，检测效率达98%。此外，还正确评估了该细胞系的突变等位基因比例，表明该技术具有定量性。