一种用于测定转染效率的流式细胞术方法。
A Flow Cytometric Method to Determine Transfection Efficiency.
作者信息
Mu Wenli, Homann Stefanie, Hofmann Christian, Gorin Alexandr, Huynh Diana, Yang Otto Orlean, Kelesidis Theodoros
机构信息
David Geffen School of Medicine, University of California, Los Angeles, USA.
Department of Medicine, Division of Infectious Diseases, David Geffen School of Medicine at UCLA, Los Angeles, California, USA.
出版信息
Bio Protoc. 2019 May 20;9(10). doi: 10.21769/bioprotoc.3244.
Mammalian cell transfection is a powerful technique commonly used in molecular biology to express exogenous DNA or RNA in cells and study gene and protein function. Although several transfection strategies have been developed, there is a wide variation with regards to transfection efficiency, cell toxicity and reproducibility. Thus, a sensitive and robust method that can optimize transfection efficiency based not only on expression of the target protein of interest but also on the uptake of the nucleic acids, can be an important tool in molecular biology. Herein, we present a simple, rapid and robust flow cytometric method that can be used as a tool to optimize transfection efficiency while overcoming limitations of prior established methods that quantify transfection efficiency.
哺乳动物细胞转染是分子生物学中常用的一种强大技术，用于在细胞中表达外源DNA或RNA并研究基因和蛋白质功能。尽管已经开发了几种转染策略，但在转染效率、细胞毒性和可重复性方面存在很大差异。因此，一种不仅可以基于目标蛋白的表达而且可以基于核酸摄取来优化转染效率的灵敏且可靠的方法，可能成为分子生物学中的重要工具。在此，我们提出一种简单、快速且可靠的流式细胞术方法，该方法可作为一种工具来优化转染效率，同时克服先前已建立的量化转染效率方法的局限性。
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