Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Cut plasmid with an appropriate restriction enzyme to produce a linear fragment for transfection. Lipofectamine transfection of c2c12 cells bridges lab. How can i optimize the transfection efficiency using lipofectamine. Lipofectamine 2000 reagent is a cationic lipidbased transfection reagent designed to give extremely high transfection efficiencies and result in equally high protein expression levels. Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene exp. Lipofectamine rnaimax transfection protocol see page 2 to view a typical rnaimax transfection procedure. Lipofectamine 2000 transfection reagent from thermo fisher. Enhanced transfection efficiency using lipofectamine 3000 reagent.
Lipofectamine 2000 transfection reagent from invitrogen. Invitrogen lipofectamine messengermax transfection reagent. Lipofectamine 2000 transfection reagent is a proprietary formulation. Highest transfection efficiency in many cell types and formats e. Lipofectamine or lipofectamine 2000 is a common transfection reagent, produced and sold by invitrogen, used in molecular and cellular biology. Lipofectamine 2000 transfection reagent is best transfection reagent available from life technologies for the co transfection. I use 24well plates for transfections in triplicates. I am performing transfection on mcf7 cells using lipofectamine 3000 invitrogen but unable to get any transfectants, instead the cells are dying after incubation with reagent and dna. Lipofectamine 2000 dna transfection reagent protocol. Lipofectamine 2000 from invitrogen biocompare product. Follow standard cloning protocol to pcr amplify the gene of interest goi and clone into the multiple cloning. Cells are dying after transfection due to toxicity of either dna lipofectamine reagent. Invitrogen cell, protein, and molecular biology technologies range from lipofectamine reagents, topo cloning kits, superscript and platinum enzymes, to western blotting technologies, antibodies, and the geneart gene synthesis service.
Lipofectamine 2000 protocol, with 2ul of lipofectamine 2000 per well. Description lipofectamine messengermax mrna transfection reagent delivers amazing transfection efficiency in neurons and a broadspectrum of primary cells, enabling improved application outcomes. Invitrogen lipofectamine rnaimax transfection reagent. Plate cells so they will be 6080% confluent at the time of transfection. Limited product warranty life technologies corporation andor its affiliates warrant their products as set forth in the life technologies general terms and conditions of sale found on life technologies. The protocol supplied by invitrogen was straight forward and also very accurate as we found that deviations from the protocol often resulted in lower transfection. Lipofectamine 2000 transfection reagent is a versatile transfection reagent that has been shown to effectively transfect the widest variety of adherent and suspension cell lines. Ideal for highthroughput work simplicity and speed combined with high transfection efficiency make lipofectamine 2000 transfection. Lipofectamine 2000 is a lipid based transfection reagent for eukaryotic cells. Scaling up or down lipofectamine 3000 reagent transfections. Shop online for a wide selection of invitrogen lipofectamine rnaimax transfection reagent provides highest transfection efficiencies on widest. Invitrogen lipofectamine 3000 transfection reagent.
According to the manufacturers protocol for 24well plate. Lipofectamine 2000 cd chemically defined transfection reagent is a proprietary animal originfree formulation for transfecting nucleic acids into eukaryotic cells. To transfect cells with sirna, follow the protocol. Lipofectamine 2000 transfection reagent is best transfection reagent available from life technologies for the co transfection of sirna and plasmid dna. Invitrogen lipofectamine 2000 transfection reagent. Lipofectamine 2000 transfection reagent is easy to usesimply mix with nucleic acid and add to cell culture. Start any new transfection by testing the recommended four concentrations of lipofectamine 2000 reagent to determine an optimum amount. As always, use good cell culture practices and wear your. Lipofectamine 2000 reagent thermo fisher scientific. Invitrogen lipofectamine 2000 transfection reagent is a proprietary formulation for the transfection of nucleic acids dna and rna into eukaryotic cells providing the following advantages. The manual recommends also that a certain amount of nucleic acid, described in ug, should be used for transfection. Hi amit, i have used optimem from invitrogen for transfecting hek 293 cells. The transfection reagent lipofectamine 2000 lf2000, invitrogen is preferred for transfection. Cells in a 6 well dish, plated and at 70% confluence.
The total volume of the transfection mixes was 100ul, and it was added to the medium already in the wells. Invitrogen lipofectamine 2000 transfection reagent 1. The product manual includes advice on how to optimize transfection efficiency, as a number of conditions can be varied including dna. This kit replaces and outperforms the original magnetofectamine which was the combination of lipofectamine 2000 from life technologies corporation and combimag transfection. A volume of 2 ul of lipofectamine 2000 do you think will be enough for an overnight transfection in dmemno pens strep 100 nm sirna.
Invitrogen cell, protein, and molecular biology technologies range from lipofectamine reagents, topo. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. What is nice about invitrogen s lipofectamine 2000 is that it has an extensive, very useful manual which addresses these issues and helps tremendously cut down the initial time of protocol. Lipofectamine 2000 transfection for hek293 cells about 6070% transfection rate transfect cells 6well plate. Plate cells so they will be 7090% confluent at the time of transfection. Exceptional transfection efficiency in the broadest range of cell lines and the highest levels of recombinant protein expression. Different cell lines were transfected with a gfp reporter vector using the indicated transfection. Achieve superior transfection efficiency while maintaining low toxicity lipofectamine 3000 reagent maintains a high transfection. Lipofectamine 3000 lipofectamine 2000 fugene hd lipofectamine 2000 lipofectamine 3000 competitor 1 competitor 2 western blot, hepg2 gststat. Invitrogen lipofectamine ltx reagent with plus reagent 0. Transfection of fluorescently labeled oligos with lipofectamine 2000 back to table of content back to protocol and application notes. Introducing the versatile reagent with the power to. Something special around the transfection protocol. The amount of lipofectamine 2000 reagent required for successful transfection varies depending on the cell type and passage number.
The alliance of mtx transfection reagent and combimag leads to increased transfection efficiency, minimized toxicity and enhanced gene expression. Prepare plasmid dnalipid complexes recommend 2 doses of lipid. Transfection p19 cells with lipofectamine 2000 the same protocol can be used for 293t cells 1. Lipofectamine 2000 cd reagent is a proprietary animal originfree. I am performing transfection on mcf7 cells using lipofectamine 3000 invitrogen but. This protocol is for transfecting plasmid dna into mammalian cells. It can be used for transfection of adherent or suspension cells with dna or rna including stealth rnai and. Lipofectamine 2000 and lipofectamine rnaimax transfection.
Protocol 1 stable transfection using lipofectamine 2000 in 6 well plates. Life technologies corporation andor its affiliates warrant their products. Using lipofectamine 2000 cd reagent for transfection provides the following advantages. Transfection of fluorescently labeled oligos with lipofectamine 2000 back to table of content back to protocol. How does transfection with lipofectamine 2000 work. Lipofectamine 2000 dna transfection reagent protocol see page 2 to view a typical dna transfection procedure. Invitrogen lipofectamine 2000 transfection reagent 0. Life technologies corporation andor its affiliates warrant their products as set. Maintain p19 cells undifferentiated in mem with 10% serum 7. For every step in your journey to discovery, there are invitrogen solutions to take you further, faster. Lipofectamine 2000 was simple to use and unlike other reagents displayed very low toxicity. Invitrogen lipofectamine 2000 transfection reagent is a proprietary. Starrlab transfection using lipoectamine transfection using lipoectamine.
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