Some cell lines are considered difficult to transfect because the correct parameters for transfection of that cell line are not known. The reagent:DNA ratio must be tested because laboratories may quantity DNA amounts slightly differently.
The Neon™ Transfection System is an improved transfection technology designed for highly efficient gene delivery into many cell types that are difficult to transfect, including primary and stem cells. The Neon™ technology uses gold-plated electrodes in a narrow, plastic pipette tip for the electroporation chamber, resulting in a more uniform electric field with minimal pH changes.
The rat pheochromocytoma PC12 cell line is widely applied as a model system to study a variety of neuronal functions. Upon addition of nerve growth factor (NGF), these cells undergo differentiation characterized by an increase in acetylcholinesterase (AChE) activity, and extension of neurite-like processes.
Lentiviruses can be used in research applications to transduce a wide range of cell types and stably integrate genetic payload into the host genome, allowing for long-term studies in vivo. This protocol makes use of ® HD Transfection Reagent and newly available components to eliminate toxicity associated with lentiviral preparations and consistently produce titers in the mid to high 1010 transduction units (TU) per milliliter.
BioTechniques Protocol Guide 2008
Some cell lines are considered difficult to transfect because the correct parameters for transfection of that cell line are not known. The reagent:DNA ratio must be tested because laboratories may quantify DNA amounts slightly differently.
BioTechniques Protocol Guide 2007
The over-expression of short interfering RNA (siRNA) in vitro is widely used to study gene functions or to identify and validate new drug targets. We describe here a highly efficient method for selective gene suppression with siRNA in various established cell lines and primary cells (full protocol available on BioTechniques Protocols-On-Line).
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