Gene Transfer
Latest Protocols
Transfection
Simple Optimization Protocol for Difficult to Transfect Cells Using FuGENE® HD Reagent
(Registration Required)
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.
Transfection
Efficient transfection of mammalian primary and stem cells using the Neon™ Transfection System
Vendor-submitted protocol. Sponsored by Invitrogen Corporation, part of Life Technologies Published in November 2009
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.
Transfection
Transfection of PC12 Cells With Lipofectamine™ LTX Does Not Affect Their Ability to Differentiate Upon NGF Stimulation
Vendor-submitted protocol. Published in November 2009
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.
Transfection
Lentiviral Production Using FuGENE® HD Transfection Reagent
Vendor-submitted protocol. Published in November 2009
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.
Transfection
Simple Optimization Protocol for Difficult to Transfect Cells Using FuGENE® HD Reagent
Vendor-submitted protocol. Sponsored by Roche Published in 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.
Electroporation
Electroporation application: Efficient selective gene suppression with siRNA.
Vendor-submitted protocol. Sponsored by Harvard Apparatus Published in 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).


Close Window