Indeed, some mirna are also known to induce gene expression by association with the promoter of the gene of interest. Maximum rna delivery is achieved with mrnain using less amount mrna than other reagents, reducing experimental costs and significantly lowering toxicity. Transfection efficiency depends on the cell type and growth rate. Hiperfect transfection reagent 1 ml from qiagen sample to. Xfect transfection reagent for plasmid transfection. It offers superior and reproducible transfection efficiency on the common cells such as hela, cho, hek293 and many hardtotransfect mammalian cells with minimal toxicity. Qiagen hiperfect transfection reagent, 1ml, cationic. Guideline of dna plasmid and hyfect tm dna transfection reagent amount and ratio refer to table 1. Increase your transfection efficiency to the level you need without killing your cells. Transitsiquest transfection reagent research reagent products. Cells may be very sensitive to the transfection reagent. Dna transfection reagent for hts applications compatible with serum and antibiotics jetpei transfection reagent is a linear polyethylenimine derivative, free of components of animal origin, providing highly effective and reproducible gene delivery.
The next day, complexes of transfection reagent and nucleic acid are added to the already adherent cells. A commonly used control is the same transfection reagent plus a noncoding rnai. It forms stable complex particles with dna and rapidly releases the dna molecules upon endocytosis into the cytoplasm. Evaluation of metafectenepro mediated transfection of sirna and. Superfect transfection reagent is provided as a readytouse solution. Im used to working with hiperfect transfection reagent for the silencing of mouse bonemarrow derived dendritic cells but want to. Purefection is a powerful, broadly applicable transfection reagent for effective and reproducible transfections.
Tpro pfect is a low toxicity, serumcompatible transfection reagent that eliminates the need for. Nanofect is a powerful, broadly applicable transfection reagent for effective and reproducible transfections. Nanofect transfection reagent is a nanotechnologybased reagent with low toxicity providing efficient gene delivery for most cell types. In order to evaluate transfection reagents under optimal conditions, test each reagent sidebyside on the same day with the same batch of cells. Use the table below to find the right product for your experiment. Transfast transfection reagent technical bulletin, tb260. Longterm gene silencing in mammalian cells using sirna eposters. In vitro dna transfection reagents from signagen laboratories. Longterm gene silencing in mammalian cells using sirna. Hiperfect transfection reagent 1 ml from qiagen sample. We are very excited to announce the introduction of our brand new product, transhi tm, as a superior transfection reagent to compete against the leading products in the market.
High transfection efficiencies and viabilities in common cell lines, particularly chinese hamster ovary cho. Altogen cro offers in vivo rnai services, tumor xenograft models, toxicology testing, stable cell line generation, and. Plate cells so they will be 6080% confluent at the time of transfection. A diverse range of transfection products is available to match the required gene delivery method. This ensures consistent transfectioncomplex formation and reproducible transfection results. To achieve effective silencing, rnai is usually applied to cells or tissue with a transfection reagent to enhance entry into cells. Acoustic liquid handling for rapid sirna transfection optimization. Lipofectamine 2000, lipofectamine rnaimax and dharmafect 3, were found to consistently give the best results. Xtremegene transfection reagents from roche have been used for a wide range of applications and a vast array of cell lines, primary cells and stem cells. Transfection reagents are highly efficient for dna and sirna transfection in vivo and in vitro. Xfect protein transfection reagent yields higher transfection efficiencies than product c across a broad range of mammalian cells, including a number of rodent and human cell lines, hardtotransfect. Transhi tm is created based on a proprietary polymer technology. Transfection using 5 nm mapk1 sirna and hiperfect transfection reagent each time cells were split resulted in prolonged knockdown of the target mrna figure 2. Multipurpose transfection reagents product information.
Pulsin powerful delivery reagent polyplustransfection. Lipofectamine rnaimax transfection protocol see page 2 to view a typical rnaimax transfection procedure. Comparison of small interfering rna sirna delivery into. Handle in a biological safety cabinet to minimize contamination. Delivery of fluorescentlylabeled sirna using transitsiquest transfection reagent. Dharmafect duo reagent for rnai molecules and plasmid delivery cotransfection of sirna or mirna reagents with a plasmid reporter system is increasingly common, yet few transfection reagents on the market will effectively deliver both molecules. What is the optimal sirna concentration for transfection of.
In addition to sirna, hiperfect transfection reagent is ideally suited to transfection of. To form complexes i incubate for 10min in 100 l optimem medium before adding. Altogen biosystems provides in vivo transfection reagents, over 100 preoptimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. Calfectin traditional recipe with new formulation most affordable and efficient best for large scale virus, antibody and protein production genjet low toxicity for very sensitive cells maximum cell viability genjet ver. In most cases, lipofectamine 2000 provides better performance than invitrogen lipofectamine transfection. In cotransfection experiments optimized for cell viability, dharmafect duo reagent resulted in. However, all the transfection reagents tested induced an ifn response in the absence of sirna, which could be minimized by reducing the transfection reagent incubation period. Compare transfection reagents transfection, electroporation. For transfection of eukaryotic cells with sirna and mirna. Hela 70% confluence cells in 12well plates were transfected with transitsiquest transfection reagent 3. Using historical comparisons transfections performed on different days to compare transfection reagents can lead to incorrect conclusions. Hiperfect transfection reagent does not need to be stored on. Interferin sirna mirna transfection reagent polyplus.
In this case it is possible to change the culture medium after 46 h. Transfection reagent, nanofect transfection reagent alstem. For use, dilute sirna in sirna dilution buffer diluted sirna is not stable, use only once. We then used a 48 well grid figure 2a to test a combination of factors, including cell density, transfection reagent, transfection reagent volume, sirna concentration, and incubation time, to determine if the adipocytes could be efficiently transfected in suspension using lipidbased reagents. Pulsin will save you time and efforts compared to other techniques using viral transduction or chemical conjugation. Transfection reagent, nanofect transfection reagent.
Pulsin reagent is readytouse and provided with a dilution buffer and a fluorescent control protein rphycoerythrin. These are available online in convenient and compact pdf. Tpro pfect transfection reagent product information. Purefection nanotechnologybased transfection reagent delivers more dna and sirnas to cells than the leading lipidbased transfection kits. Tpro pfect transfection reagent is a broad spectrum, low toxicity transfection reagent that provides high efficiency plasmid dna delivery in many mammalian cell types. In co transfection experiments optimized for cell viability, dharmafect duo reagent resulted in. The transfast reagent is supplied as a dried lipid film that forms multilamellar vesicles upon hydration with water. Nanofect transfection reagent is a unique blend of polymers that delivers more dna and sirnas to cells than the leading lipidbased transfection kits, at a lower cost. Transfection is a welltested and crucial biotechnology that allows researchers to study gene products and gene functioning directly in cells.
Viafect transfection reagent is a cationic delivery reagent in aqueous solution. In the twostep transfection method, the cells are plated 24 hours before transfection. Superfect transfection reagent from qiagen selectscience. Qiagen hiperfect transfection reagent, 1ml, cationic and neutral lipids technology, transient transfection, eukaryotic cells including primary cells type, minimized risk of offtarget effects, rapid reverse transfection, for up to 333 transfections in 24well plates or up to 33. Hiperfect transfection reagent 1 ml, 301705 from qiagen. The polyjet reagent is able to immobilize dna migration during.
The examples given within the protocol are for 96well plates, and table 1 provides the transfection reagent volumes for additional plate types. Sep 11, 2009 we then used a 48 well grid figure 2a to test a combination of factors, including cell density, transfection reagent, transfection reagent volume, sirna concentration, and incubation time, to determine if the adipocytes could be efficiently transfected in suspension using lipidbased reagents. In addition to basic transfection kits, enhancement compounds can be used to increase the efficiency of the selected transfection treatment. Data below also shows mrnain significantly outperforms competitor reagents, typically achieving 90% or greater transfection efficiency across a wide range of cell types.
Im used to working with hiperfect transfection reagent for the silencing of mouse bonemarrow derived. The efficacy of eleven transfection reagents to promote uptake of the fitclabelled sirna was investigated. When cells were transfected only once, repeated splitting cycles resulted in loss of the silencing effect figure 2. The reagent is simply added to the dna solution, mixed, and incubated for 510 minutes to allow superfectdna complexes to.
Xfect is a biodegradable transfection polymer, screened from a pool of 2,300 candidates for its very low cytotoxicity profile and high transfection efficiency. This allows the use of transfected cells in multiday assays where overly con. Ii 100% dna condensing groups released efficient for commonly used cell types genjet. Hiperfect transfection reagent qiagen for sirna transfection in adherent cells. The purefection reagent selfassembles nanoparticles in the presence of dna and rna. Viafect transfection reagenta is a novel formulation designed to transfect dna into a wide variety of cell lines with high efficiency and low.
Nonlipid based chemical transfection reagent optimized for maximum transfection efficiency, easeofuse, and minimal cytotoxicity on a wide variety of mammallian cells. Lipofectamine ltx, lipofectamine rnaimax, and hiperfect were chosen for the next. In vivo transfection reagents are offered with markers, such as neomycin resistance. Hiperfect transfection reagent 1 ml from qiagen selectscience. Xfect transfection reagent for plasmid transfection increase your transfection efficiency to the level you need without killing your cells. Transitsiquest transfection reagent research reagent. What is the optimal sirna concentration for transfection. Lipofectamine rnaimax reagent thermo fisher scientific. Protein transfection efficiencies across different cell lines. Dissolve sirna in sterile water and prepare 5 l aliquots, store at 80c.
Total volume of medium, dna and viafect transfection reagent for 96well plates at a 3. Dharmafect duo reagent for rnai molecules and plasmid delivery co transfection of sirna or mirna reagents with a plasmid reporter system is increasingly common, yet few transfection reagents on the market will effectively deliver both molecules. The cells were incubated 24 hours posttransfection then fixed and counterstained with. The transfast reagent is supplied as a dried lipid film that forms. Rna interference rnai is a powerful platform utilized to target transcription of specific genes and downregulate the protein product. Purefection nanotechnologybased transfection reagent. Transfections can be performed in the presence of serum, eliminating the need to remove complexes from the cells. Viafecttm transfection reagent technical manual tm409. Hiperfect transfection reagent or qiagen products in general, please do not hesitate to contact. Viafect transfection reagent typically demonstrates low toxicity at low cell densities and therefore can be used on cells at low con.
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