Rose John K
Yale University School of Medicine, New Haven, Connecticut, USA.
Curr Protoc Cell Biol. 2003 Aug;Chapter 20:Unit 20.7. doi: 10.1002/0471143030.cb2007s19.
When embarking upon any transfection procedure, a critical first step is to optimize conditions. Every mammalian cell type has a characteristic set of requirements for optimal introduction of foreign DNA; there is a tremendous degree of variability in the transfection conditions that work, even among cell types that are very similar to one another. Often, an experimenter must screen a wide variety of cell types for a desired regulatory trait, such as an appropriate response to a particular effector molecule. It is thus helpful to have a straightforward, systematic approach to optimizing transfection efficiency. This unit provides guidelines to optimizing transfection experiments.
在开始任何转染程序时,关键的第一步是优化条件。每种哺乳动物细胞类型对于外源DNA的最佳导入都有一套特定的要求;即使在彼此非常相似的细胞类型之间,有效的转染条件也存在极大的差异。通常,实验者必须针对所需的调控特性(例如对特定效应分子的适当反应)筛选多种细胞类型。因此,采用一种直接、系统的方法来优化转染效率会很有帮助。本单元提供了优化转染实验的指南。