Compare transfection reagents transfection, electroporation. Description based on our proprietary polymer synthesis technology, polyjet dna in vitro transfection reagent is formulated as a biodegradable polymer based dna transfection reagent that ensures effective and reproducible transfection on hek293, cos7, nih3t3, hela, cho and a broad ranges of hardtotransfect mammalian cells. 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. In vitro dna transfection reagents from signagen laboratories. Longterm gene silencing in mammalian cells using sirna. Lipofectamine ltx, lipofectamine rnaimax, and hiperfect were chosen for the next. Rna interference rnai is a powerful platform utilized to target transcription of specific genes and downregulate the protein product. In this case it is possible to change the culture medium after 46 h. 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. Superfect transfection reagent from qiagen selectscience. Indeed, some mirna are also known to induce gene expression by association with the promoter of the gene of interest. Pulsin will save you time and efforts compared to other techniques using viral transduction or chemical conjugation.
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. 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. 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. The purefection reagent selfassembles nanoparticles in the presence of dna and rna. Xfect transfection reagent for plasmid transfection. In addition to sirna, hiperfect transfection reagent is ideally suited to transfection of. Delivery of fluorescentlylabeled sirna using transitsiquest transfection reagent. For use, dilute sirna in sirna dilution buffer diluted sirna is not stable, use only once. Maximum rna delivery is achieved with mrnain using less amount mrna than other reagents, reducing experimental costs and significantly lowering toxicity. Hiperfect transfection reagent 1 ml from qiagen sample to. Lipofectamine rnaimax transfection protocol see page 2 to view a typical rnaimax transfection procedure.
Purefection nanotechnologybased transfection reagent. 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. Longterm gene silencing in mammalian cells using sirna eposters. In co transfection experiments optimized for cell viability, dharmafect duo reagent resulted in. Increase your transfection efficiency to the level you need without killing your cells. 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. Use the table below to find the right product for your experiment. The next day, complexes of transfection reagent and nucleic acid are added to the already adherent cells. Superfect transfection reagent is provided as a readytouse solution. Dissolve sirna in sterile water and prepare 5 l aliquots, store at 80c. Nanofect is a powerful, broadly applicable transfection reagent for effective and reproducible transfections. 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 efficiency lipidbased sirna transfection of adipocytes. Im used to working with hiperfect transfection reagent for the silencing of mouse bonemarrow derived. Qiagen hiperfect transfection reagent, 1ml, cationic. Transitsiquest transfection reagent research reagent. For transfection of eukaryotic cells with sirna and mirna. This allows the use of transfected cells in multiday assays where overly con. Tpro pfect transfection reagent is a broad spectrum, low toxicity transfection reagent that provides high efficiency plasmid dna delivery in many mammalian cell types. High transfection efficiencies and viabilities in common cell lines, particularly chinese hamster ovary cho. In most cases, lipofectamine 2000 provides better performance than invitrogen lipofectamine transfection.
In brief, plasmid and transfection reagent are diluted in serumfree culture medium for 5 mins then mixed together and tip gently for another 25 mins. Xfect is a biodegradable transfection polymer, screened from a pool of 2,300 candidates for its very low cytotoxicity profile and high transfection efficiency. What is the optimal sirna concentration for transfection of. Viafect transfection reagenta is a novel formulation designed to transfect dna into a wide variety of cell lines with high efficiency and low. Ii 100% dna condensing groups released efficient for commonly used cell types genjet.
I do recommend testing different sirna concentrations and hiperfect amounts for your particular cell line. The examples given within the protocol are for 96well plates, and table 1 provides the transfection reagent volumes for additional plate types. Purefection is a powerful, broadly applicable transfection reagent for effective and reproducible transfections. Transfection reagent, nanofect transfection reagent. Transitsiquest transfection reagent research reagent products. The transfast reagent is supplied as a dried lipid film that forms multilamellar vesicles upon hydration with water. Hiperfect transfection reagent 1 ml, 301705 from qiagen. Pulsin reagent is readytouse and provided with a dilution buffer and a fluorescent control protein rphycoerythrin.
Pulsin powerful delivery reagent polyplustransfection. Nonlipid based chemical transfection reagent optimized for maximum transfection efficiency, easeofuse, and minimal cytotoxicity on a wide variety of mammallian cells. Plate cells so they will be 6080% confluent at the time of transfection. Hiperfect transfection reagent is provided as a readytouse solution just add the reagent to your diluted sirnamirna, mix, incubate, and pipet the complexes onto the cells. Hiperfect transfection reagent 1 ml from qiagen sample. Transhi tm is created based on a proprietary polymer technology. Place 5000 cells in 175 ul of complete media and add to 96 well containing the sirna and hiperfect. Transfection reagent, nanofect transfection reagent alstem.
Tpro pfect is a low toxicity, serumcompatible transfection reagent that eliminates the need for. Total volume of medium, dna and viafect transfection reagent for 96well plates at a 3. Transfection efficiency depends on the cell type and growth rate. In cotransfection experiments optimized for cell viability, dharmafect duo reagent resulted in. Transfection is a welltested and crucial biotechnology that allows researchers to study gene products and gene functioning directly in cells. Xfect transfection reagent for plasmid transfection increase your transfection efficiency to the level you need without killing your cells. Evaluation of metafectenepro mediated transfection of sirna and. These are available online in convenient and compact pdf. Cells may be very sensitive to the transfection reagent. This ensures consistent transfectioncomplex formation and reproducible transfection results. The transfast reagent is supplied as a dried lipid film that forms. 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. Protein transfection efficiencies across different cell lines. 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.
Altogen cro offers in vivo rnai services, tumor xenograft models, toxicology testing, stable cell line generation, and. Lipofectamine rnaimax reagent thermo fisher scientific. Nanofect transfection reagent is a nanotechnologybased reagent with low toxicity providing efficient gene delivery for most cell types. Hiperfect transfection reagent or qiagen products in general, please do not hesitate to contact. When cells were transfected only once, repeated splitting cycles resulted in loss of the silencing effect figure 2.
To form complexes i incubate for 10min in 100 l optimem medium before adding. Viafecttm transfection reagent technical manual tm409. It forms stable complex particles with dna and rapidly releases the dna molecules upon endocytosis into the cytoplasm. A commonly used control is the same transfection reagent plus a noncoding rnai. Transfection reagents are highly efficient for dna and sirna transfection in vivo and in vitro. Tpro pfect transfection reagent product information. 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. Hiperfect transfection reagent 1 ml from qiagen selectscience.
Viafect transfection reagent is a cationic delivery reagent in aqueous solution. The polyjet reagent is able to immobilize dna migration during. Comparison of small interfering rna sirna delivery into. Guideline of dna plasmid and hyfect tm dna transfection reagent amount and ratio refer to table 1. In addition to basic transfection kits, enhancement compounds can be used to increase the efficiency of the selected transfection treatment. Hiperfect transfection reagent does not need to be stored on. Handle in a biological safety cabinet to minimize contamination. Hiperfect transfection reagent qiagen for sirna transfection in adherent cells. Using historical comparisons transfections performed on different days to compare transfection reagents can lead to incorrect conclusions.
Transfections can be performed in the presence of serum, eliminating the need to remove complexes from the cells. 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. Altogen biosystems provides in vivo transfection reagents, over 100 preoptimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. In the twostep transfection method, the cells are plated 24 hours before transfection. Hela 70% confluence cells in 12well plates were transfected with transitsiquest transfection reagent 3. Viafect transfection reagent typically demonstrates low toxicity at low cell densities and therefore can be used on cells at low con. Purefection nanotechnologybased transfection reagent delivers more dna and sirnas to cells than the leading lipidbased transfection kits.
The reagent is simply added to the dna solution, mixed, and incubated for 510 minutes to allow superfectdna complexes to. Im used to working with hiperfect transfection reagent for the silencing of mouse bonemarrow derived dendritic cells but want to. In order to evaluate transfection reagents under optimal conditions, test each reagent sidebyside on the same day with the same batch of cells. What is the optimal sirna concentration for transfection. The cells were incubated 24 hours posttransfection then fixed and counterstained with. The efficacy of eleven transfection reagents to promote uptake of the fitclabelled sirna was investigated. Incubate at rt 5 minutes i have waited as long as 30 minutes. In vivo transfection reagents are offered with markers, such as neomycin resistance. First generation reagent for plasmid dna transfections. It offers superior and reproducible transfection efficiency on the common cells such as hela, cho, hek293 and many hardtotransfect mammalian cells with minimal toxicity. Multipurpose transfection reagents product information.
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