使用Lipofectamine 2000试剂进行高级转染：原代神经元、小干扰RNA及高通量应用。

Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications.

作者信息

Dalby Brian, Cates Sharon, Harris Adam, Ohki Elise C, Tilkins Mary L, Price Paul J, Ciccarone Valentina C

机构信息

Invitrogen Corporation Research and Development, 1610 Faraday Avenue, Carlsbad, CA 92008, USA.

出版信息

Methods. 2004 Jun;33(2):95-103. doi: 10.1016/j.ymeth.2003.11.023.

DOI:10.1016/j.ymeth.2003.11.023

PMID:15121163

Abstract

Lipofectamine 2000 is a cationic liposome based reagent that provides high transfection efficiency and high levels of transgene expression in a range of mammalian cell types in vitro using a simple protocol. Optimum transfection efficiency and subsequent cell viability depend on a number of experimental variables such as cell density, liposome and DNA concentrations, liposome-DNA complexing time, and the presence or absence of media components such as antibiotics and serum. The importance of these factors in Lipofectamine 2000 mediated transfection will be discussed together with some specific applications: transfection of primary neurons, high throughput transfection, and delivery of small interfering RNAs.

摘要

Lipofectamine 2000是一种基于阳离子脂质体的试剂，通过简单的方案在体外多种哺乳动物细胞类型中提供高转染效率和高水平的转基因表达。最佳转染效率及随后的细胞活力取决于许多实验变量，如细胞密度、脂质体和DNA浓度、脂质体 - DNA复合时间，以及培养基成分（如抗生素和血清）的存在与否。将讨论这些因素在Lipofectamine 2000介导的转染中的重要性以及一些具体应用：原代神经元的转染、高通量转染和小干扰RNA的递送。