This study shows that lipofectamine 2000 is an efficient reagent for the delivery of genes into sw480 cells. For the cell lines easy to transfection, you can use either of the procedures reverse or forward. Theres a difference between the two protocols lipofectamine 3000 has an extra reagent that needs to be added. Jul 17, 2007 lipofectamine 2000 is a lipid based transfection reagent for eukaryotic cells. The amount of lipofectamine 2000 reagent required for successful transfection varies depending on the cell type and passage number. Mix optimem and lipofectamine in another eppendorf 32 l lipofectamine in 0,8 ml optimem 3. Comparison of k2 transfection system with lipofectamine.
Enable high transfection efficiency in novel genome editing applications. This is most often an endogenous proteincoding gene, although some noncoding rnas can also be silenced effectively. Use a range of several ul of lipofectamine and ug dna to increase viability after transfection and efficiency. At the foundation of the popularity of this broadspectrum reagent is the reliably high efficiency and simple protocol for success across a broad range of cell lines figure 1. Lipofectamine 2000 is used to deliver the sirnas into hek293 cells. Lipofectamine 2000 transfection reagent is best transfection reagent available from life technologies for the cotransfection of sirna and plasmid dna. Researchers use lipofectamine 2000 reagent for sirna and shrnabased gene knockdown experiments, as well as for gene expression studies. Make dnalipofectamine 3000 complexes in serumfree medium such as. Hi, i was wondering if someone know the optimal lipofectamine 2000 to dna ratio for transfecting saos2 cells. Comparison of k2 transfection system with lipofectamine 2000 invitrogen for the transient transfection of hek293t cells. Add 100l of optimum to the eppendorf tube to dilute the dna and mix by tapping. Highest transfection efficiency in many cell types and formats e. Lipofection is most widely used method for transfecting neurons for livecell imaging experiments.
Add 6l of lipofectamine2000 from biostores and 100 l of. Transfection efficiency in a cancer cell line panel. Add diluted dna to each tube of diluted lipofectamine 2000 reagent 1. Prewarm 50ml of optimum stored in cold room at 4c 10min. Invitrogen lipofectamine 3000 transfection reagent fisher. Step tube complexation components amount per well 24well 1 tube 1 optimem i medium 25 l lipofectamine 3000 reagent 1. Lipofectamine 2000 reagent and lipofectamine ltx reagent. In order to minimize the adverse environmental impact of packaging and shipping products on gel ice, we investigated the feasibility of shipping four of. Start any new transfection by testing the recommended four concentrations of lipofectamine 2000 reagent to determine an optimum amount. The lipofectamine rnaimax reagent is designed specifically for the delivery of sirna and mirna, and lipofectamine messengermax reagent delivers mrna. However, for cotransfection of sirnachimera and plasmid dna, lipofectamine 2000 should be used. Lipofectamine 3000 transfection reagent leverages our most advanced lipid nanoparticle technology to provide superior transfection performance with improved application outcomes and reproducible results. Pdf optimization of sw480 colon cancer cells transfection. Lipofectamine 2000 reagent thermo fisher scientific.
Lipofectamine 2000 transfection for hek293 cells about 6070% transfection rate transfect cells 6well plate. Transfection of nih3t3 cells, hela, swis 3t3, 293t with lipofectamine 2000 1. The trusted, simple, broadspectrum transfection reagent for most cell lines. Mix optimem with dna in an eppendorf 510 g dna0,8 ml optimem 2. 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.
Lipofectamine 3000 transfection reagent hepg2 liver. The protocal is from thermo fisher scientific important guidelines for transfection 1. Lipofectamine rnaimax transfection reagent is a proprietary formulation for transfecting small rnas \e. Transfection commonly refers to the introduction of nucleic acids into eukaryotic cells, or more specifically, into animal cells. I personally have used this reagent to transfect cells for fluorescent microscopy, gst pull downs, and coimmunoprecipitation experiments. Rnai in cultured mammalian cells using synthetic sirnas. Lipofectamine 2000 reagent protocol 202lipofectamine 2000 dna transfection reagent protocol transfect cells according to the following chart. Lipofectamine 2000 reagent delivers dna or sirna with excellent transfection performance for protein expression, gene silencing, and functional assays. It can be used for transfection of adherent or suspension cells with dna or rna including stealth rnai and sirna and can be used for either transient transfections or to create stable cell lines. Being a lowefficiency method it results in very sparse distribution of transfected neurons, which is perfect for analysis of endosomes dynamics within single neurons. Lipofectamine 2000 transfection reagent from invitrogen. Invitrogen lipofectamine 3000 transfection reagent from.
Lipofectamine 2000 and lipofectamine rnaimax transfection reagents introduction life technologies is committed to providing researchers and scientists the best products, enabling cutting edge technology while keeping the environment in mind. Versatileone reagent for dna, rna, and cotransfection. Transfection of nih3t3 cells, hela, swis 3t3, 293t with. Transfecting sirna using lipofectamine raiax pin liu, 2017 declaration. Invitrogen lipofectamine 2000 transfection reagent fisher. Lipofectamine 2000 is a lipid based transfection reagent for eukaryotic cells.
Effi ciency and ofp expression were analyzed 72 hours posttransfection and a u2os and b hepg2 cells showed 4fold and 80fold improvement with lipofectamine. Transfection protocol lipofectamine 3000 transfection reagent. Evaluation of transfection methods for transient gene. Msc victor van puyenbroeck, rega institute for medical research, department of microbiology and immunology, university of leuven, leuven, belgium. I use 24well plates for transfections in triplicates. Lipofectamine 2000 reagent dna seed cells so they are 7090% con. Lipofectamine 2000 and 3000 summary in this comparative study, transfections were performed with dnain and compared to the current market leader, lipofectamine 2000 lf2k, using a series of cultured celllines and primary cells. The vector contained an ofp reporter gene and was transfected with lipofectamine 2000 or lipofectamine 3000 reagent into a u2os and b hepg2 cell lines.
Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. Lipofectamine reagents go online to view related products. Lipofectamine 2000, and lipofectamine 2000 cd x broad spectrum dna deliveryachieve high expression in many cell types, including hardtotransfect cells and insect cells x animal origin freetransit2020 provides high performance with maximum compatibility description transit2020 reagent is a new and versatile transfection solution for. Lipofectamine 3000 reagent is a proprietary formulation for transfecting nucleic acids into a wide range of eukaryotic cells and especially designed for hard to transfect cells. Transfection protocol of lipofectamine rnaimax can be obtained from the invitrogen website.
Lipofectamine 2000 reagent and lipofectamine 3000 reagent were used to transfect 17 cell lines with a gfpexpressing plasmid in a 24well plate format using 0. Lipofectamine 2000 transfection reagent is versatile transfection reagent that has been shown to effectively transfect widest variety of adherent and suspension cell lines. Lipofectamine 2000 and lipofectamine 3000 reagents deliver dna or cotransfect with excellent transfection performance for protein expression, gene silencing, and functional assays. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6well transfections, and accounts for pipetting variations. 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.
We have had success using lipofectamine reagents in the past and the lipofectamine 3000 appears to be the best option. Lipofectamine 2000 transfection protocol 10cm plates reagents. For your convenience, the essential components of this protocol are now available in the gibco liver cancer starter kit. Lipofectamine 2000 and lipofectamine rnaimax transfection. Lipofectamine 2000 transfection reagent is a proprietary formulation for the transfection of nucleic acids dna and rna into eukaryotic cells and provides the following advantages. Formulation of lipofectamine 2000 and lipofectin signagen blog. Invitrogen lipofectamine 2000 transfection reagent. The diluted dna and lipofectamine 2000 were combined and incubated at. Make sufficient 10% fbs in optimem media wo antibiotics sterile filter 2. I use lipofectamine 2000 invitrogen for transfection of both sirna and plasmid dna into these 293 cell lines. Invitrogen lipofectamine 2000 transfection reagent is a versatile transfection reagent that has been shown to effectively transfect a variety of payloads into the widest variety of adherent. Lipofectamine 2000, 3000, messengermax, and rnaimax. Photograph of expected results dna lipofectamine scaling up or down. Dilute 4 amounts of lipofectamine 2000 reagent in optimem medium.
Optimum amount needed is determined from the protocol for lipofectamine 3000 transfection reagent. Efficiently transfect difficulttotransfect cells lipofectamine 3000 reagent is designed to efficiently transfect difficulttotransfect cells, yielding superior transfection performance across the broadest array of. Invitrogen lipofectamine 2000 transfection reagent 1. Refer to the cell lines database for a list of cell types successfully. Invitrogen lipofectamine 3000 transfection reagent. Transfection and genome engineering fisher scientific. Transfection protocol lipofectamine 3000 transfection. Invitrogen lipofectamine 2000 transfection reagent strong performance in a wide range of cell lines reliableone of the mostcited dna reagents strong performancefor easytotransfect to moderately difficulttotransfect cell lines easy to usesimple protocol for easy optimization in a wide range of cells. The lipofectamine 3000 is the bees knees though, i managed to get loads of transfects from a difficult to transfect cell line that had been giving us trouble with the lipofectamine 2000. Lipofectamine 2000 transfection reagent from thermo fisher.