The product manual includes advice on how to optimize transfection. Transfection efficiencies using commercially available reagents such as fugene hd and lipofectamine 2000 were generally low, but even the best. Use the procedure on page 3 to transfect cells with plasmid dna. Use this brief procedure to transfect stealth rnai or sirna into mammalian cells. Reverse transfection an overview sciencedirect topics. Lipofectamine 2000 is commonly used for sirna transfections. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6well transfections, and accounts for pipetting variations. Lipofectamine 2000 and lipofectamine rnaimax transfection. Effects of lipofectamine 2000sirna complexes on autophagy in. Lipofectamine 2000 is a commonly used reagent for sirna delivery. Guidelines for transfection use the procedure on below to transfect cells with short interfering rna sirna or invitrogen stealth rnai. Transfection protocol lipofectamine 3000 transfection. Stealthsirna transfection protocol lipofectamine 2000. Lipofectamine rnaimax transfection protocol see page 2 to view a typical rnaimax transfection procedure.
Optimized delivery of sirna into 3d tumor spheroid. Can i use lipofectamine 2000 to transfect with sirna. Lipofectamine 2000 dna transfection reagent protocol. To transfect cells with sirna, follow the protocol as described for dna but. Invitrogen lipofectamine 2000 transfection reagent is a proprietary formulation. Lipofectamine 2000 transfection reagent from invitrogen. Systematic screening of commonly used commercial transfection. Looking to do some preliminary experiments and i dont want to but a new reagent if i dont have to.
So i assumed this system was pretty forgiving in terms of following instructions. I transfect my pc3 cell line with lipofectamine 2000 and my sirna plk1 and with. Transfecting sirna using lipofectamine rnaimax pin. For each reagent, the manufacturers own instructions should be followed.
Lipofectamine 2000 life technologies or similar transfection reagent. Lipofectamine 2000 reagent thermo fisher scientific. Follow these general guidelines when using lipofectamine 2000 to cotransfect your plasmid dna and the rnai molecule of interest into mammalian cells. Co transfection of plasmid dna and sirna transfect plasmid dna and sirna at the same time using lipofectamine 2000 reagent by adding 30 pmol 0. How does transfection with lipofectamine 2000 work. Rnaimax, a reagent designed for double stranded sirna delivery. In the protocol described here, we use a pool of three sirna duplexes per target gene per well, whereas, for example, using three duplexes per. Lipofectamine 2000 protocol, with 2ul of lipofectamine 2000 per well. Ca using lipofectamine 2000 invitrogen according to manufacturers instructions. Transfect plasmid dna and sirna at the same time using lipofectamine 2000. Use the procedure on below to transfect cells with plasmid dna. Use the procedure on page 2 to transfect cells with short interfering rna sirna or invitrogen stealth rnai.