Description:
The TransPass™ HeLaTransfection Reagent is a lipid/cationic polymer formulation that can be used for efficient transfection of plasmid DNA or siRNA into HeLa cells with a serum-compatible protocol. Co-transfection of plasmid and siRNA can also be efficiently performed.
Background:
The introduction of recombinant DNA into cultured cells or "transfection", has become an essential tool for studying gene function and regulation. Initially, cells were transfected by methods such as Calcium Phosphate co-precipitation, electroporation or viral factors (1, 2). The introduction of cationic lipids or polymers as transfection agents has led to the development of reagents that efficiently deliver DNA through the cell membrane and into the nucleus (3). However, not all DNA transfection reagents can deliver siRNA as efficiently as plasmid DNA and vice versa.
Transfection Guidelines:
For consistent results, it is important to maintain healthy proliferating cells that are regularly passaged.
It is important that NO heparin and NO antibiotics/antimycotics in the growth medium during transfection.
Use sterile plasmid DNA that is purified by CsCl gradient centrifugation or column chromatography.
The following parameters can be optimized in order to maximize the transfection efficiency for a particular cell line: the cell density at the time of transfection, amount of transfection reagent, amount of plasmid DNA and the culture incubation time before analysis. The specific amount of transfection reagent can be optimized for a particular cell line by performing a simple titration
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