Xtremegene sirna transfection reagent sigmaaldrich. Optimization of sirna transfection for breast and glioma. Roche applied science has recently introduced the x tremegene sirna transfection reagent according to the company, the reagent is an optimized formulation that forms a complex with sirna, as well as with mixtures of sirna and plasmid dna. Xtremegene sirna transfection reagent polymer reagent for. Transfection reagents are highly efficient for dna and sirna transfection in vivo and in vitro. Transfection protocol lipofectamine 3000 transfection. Need xtremegene hp dna transfection reagent for research. Small interfering rna sirna highthroughput screening has a variety of. Xfect is a transfection reagent that creates biodegradable nanoparticles that permit superior transfection efficiency of plasmid dna into mammalian cells. Jun 30, 2016 reliable and efficient dna and rna transfection methods are required when studying the role of individual genes in mouse pluripotent stem cells. Shared reagents dna transfection sirna transfection growth medium optimem medium for complexing dna p3000 reagent lipofectamine 3000 reagent sirna lipofectamine 3000 reagent 96well 0.
This transfection reagent panel was assembled by the target discovery institute hts facility as a representative sample of the sirna transfection reagents currently available. Last time i did transfection using roche xtremegene sirna transfection reagent during second day of differentiation media stage. With our proprietary ph dependent conformational change pdcc technology, the biodegradable transfection reagent was formulated by addition of prescreened hydrophobic groups to its side chain, making sitran 2. Metafectene is suitable for transient and stable transfection of mammalian cell lines with dna plasmids, bacmids, rna mrna, mirna and sirna and modified nucleic acids antisense oligonucleotides. Roche applied science s xtremegene sirna transfection reagent. Unlike for conventional dna transfection, transport of the nucleic acid into the cell. Xtremegene hp dna transfection reagent highperformance. Fast and efficient transfection of mouse embryonic stem. Xtremegene hp dna transfection reagent highperformance polymer reagent for transfecting many cell lines. In addition to basic transfection kits, enhancement compounds can be used to increase the efficiency of the selected transfection treatment.
X tremegene 9 transfection reagent 3 l 15 l dna 1 g 5 g 6. Hiperfect transfection reagent handbook 102010 7 safety information when working with chemicals, always wear a suitable lab coat, disposable gloves, and protective goggles. Xtremegene 9 dna transfection reagent polymer reagent. General description short interfering rna sirna can be directly introduced into cells by transfection. Optimization of sirna transfection for breast and glioa. Altogen biosystems provides in vivo transfection reagents, over 100 preoptimized in vitro transfection kits for cell lines and primary cells, and electroporation delivery products. The xtremegene sirna transfection reagent is intended for research applications only. Delivery of fluorescentlylabeled sirna using transittko transfection reagent. The reagent is free of any animalderived components. Xtremegene dna xtremegene dna transfection reagent. The x tremegene sirna transfection reagent delivers all the benefits of high transfection efficiency, low cytotoxicity, and the capability of transfecting a.
On the day before transfection, plate cells so they will be 5070% confluent on. Find transfection solutions for efficient delivery of dna, sirna, oligos, and rna into adherent and suspension cells, including hardtotransfect cells. For the most current list of successfully transfected cell types, visit the x. Trans it transfection reagents are designed, tested and manufactured in madison, wi. High efficiency versatile reagent for a wide range of common cell types most efficient versatile reagent for the widest range of cell types including difficulttotransfect cells most efficient reagent for. We support multiple transfection methods with a different reagent for nearly any application. Comparison of small interfering rna sirna delivery into bovine monocytederived macrophages by transfection and electroporation.
In addition to basic transfection kits, enhancement compounds can be used to increase the efficiency of. Roche applied science has recently introduced the xtremegene sirna transfection reagent according to the company, the reagent is an optimized formulation that forms a complex. Invitrogen lipofectamine 2000 transfection reagent. Culture vessel multiplication factor shared reagents dna transfection sirna transfection growth medium optimem medium for complexing dna p3000 reagent lipofectamine 3000 reagent sirna lipofectamine 3000 reagent. Optimizing ratio of x tremegene sirna transfection reagent, sirna, and plasmid dna in various plate formats refer to table 1 when setting up your transfection reactions in other plate formats. These properties make sirna transfection a useful tool for gene silencing in mammalian cells. A diverse range of transfection products is available to match the required gene delivery method. Dna and sirna transfection reagent, jetprime transfection reagents the jetprime dna andor sirna transfection reagent is a powerful and versatile reagent designed to ensure high dna transfection efficiency and excellent gene silencing in a variety of adherent cells. Xtremegene sirna transfection reagent from roche applied.
A lipidbased transfection reagent, such as xtremegene sirna transfection reagent, can provide a convenient, reliable, and efficient vehicle for delivering sirnas into animal cells, enabling the study of cellular and functional consequences of gene knockdown. With our proprietary ph dependent conformational change pdcc. Xtremegene hp dna transfection reagent is a high performance reagent that transfects a broad range of eukaryotic cells, including insect cells and hardtotransfect cell lines e. A detailed protocol is available in the online instructions for use at structions. Transfections can be carried out entirely in the presence of serum. Our reagents are ideal for delivering multiple types of nucleic acids including. Such a reagent is x tremegene sirna transfection reagent, which forms a complex with sirna and effectively delivers it into eukaryotic cells. Pepmute sirna transfection reagent, formulated from simulation of virus cell penetrating peptides cpps, is a total novel sirna delivery tool which provides more than 95% silencing efficiency at 1 nm sirna in variety of mammalian cells. Xtremegene 9 transfection reagent 3 l 15 l dna 1 g 5 g 6. U87 mg transfection results with silentfect and xtremegene. Locate proteins, assay kits, reagents, custom services.
These are available online in convenient and compact pdf. Invitrogen lipofectamine rnaimax transfection reagent. Xtremegene sirna transfection reagent polymer reagent. With our proprietary ph dependent conformational change pdcc technology, the. Transfection protocol lipofectamine 3000 transfection reagent. A lipidbased transfection reagent, such as xtremegene sirna transfection reagent, can provide a. Antibiotics will inhibit transfection complex formation and therefore should be. Find and compare commercial and governmental sources for immunological and biological products using the linscotts directory search. Efficient delivery of dna, sirna, mirna, and crisprcas9 components with a joint commitment to enable your next discovery, we exclusively offer the complete portfolio of roche x tremegene transfection reagents. Reliable and efficient dna and rna transfection methods are required when studying the role of individual genes in mouse pluripotent stem cells. For the most current list of successfully transfected cell types, visit. In vivo transfection reagents are offered with markers, such as neomycin resistance.
Control duplex using xtremegene roche transfection reagent. In contrast, xtremegene and lipofectamine 2000 were significantly lower. Rnaishuttle sirna transfection reagent is developed to allow researchers to fully take. The jetprime dna andor sirna transfection reagent is a powerful and versatile reagent designed to ensure high dna transfection efficiency and excellent gene silencing in a variety of adherent cells. Tips and tricks on how to improve transfection experiments. The cytocompatibility of the vectorsirna polyplexes was assessed by viability assay of cells.
Xtremegene 9 dna transfection reagent sigmaaldrich. Interferin sirna mirna transfection reagent polyplus. Indeed, some mirna are also known to induce gene expression by association with the promoter of the gene of interest. Optimization of sirna transfection for breast and glioma cancer cell cultures rna silencing technology and methods based thereupon are important in todays medical biotechnology. Altogen cro offers in vivo rnai services, tumor xenograft models, toxicology testing, stable cell line generation, and.
Lipofectamine rnaimax exhibited much higher sirna transfection efficiency than the other reagents, and fugene hd was identified as the best suitable reagent for efficient pdna transfection of hes. On the day of transfection, which should be 1 day following cell plating, perform the following steps, which have been optimized for a single well of a 24well plate using lipofectamine 3000 transfection. Fast and efficient transfection of mouse embryonic stem cells. Xx easytofollow procedure, for any cell type hiperfect transfection reagent is provided as a readytouse solution just add the reagent to your diluted. Rnaishuttle sirna transfection reagent instruction manual. With our proprietary ph dependent conformational change pdcc technology, the biodegradable transfection reagent was formulated by addition of prescreened hydrophobic groups to its side chain, making genmute reagent a versatile and most powerful gene delivery tool in the market. On the day before transfection, plate cells so they will be 5070% confluent on transfection day.
As a consequence of endosome buffering nucleic acids are released more easily to the cytosol, thus increasing the efficiency of transfection. The levels of sirna uptake were observed to increase with. We support multiple transfection methods with a different reagent for nearly any. Fullydetailed protocols are available on product pages.
The starting volume and mass is based on a x tremegene sirna transfection reagent. Efficient sirna delivery and gene silencing using a lipopolypeptide. Hela 70% confluence cells in 12well plates were transfected with transittko transfection reagent 3. Use the following table to scale the volumes for your transfection experiment. Culture vessel multiplication factor shared reagents dna transfection sirna. Pepmute sirna transfection reagent, formulated from simulation of virus cell penetrating peptides cpps, is a total novel sirna delivery tool which provides more than 95% silencing efficiency. Comparison of small interfering rna sirna delivery into. Optimizing ratio of xtremegene sirna transfection reagent, sirna, and plasmid dna in various plate formats refer to table 1 when setting up your transfection reactions in other plate formats. Invitrogen lipofectamine rnaimax transfection reagent 1. Cd vector was evaluated by flow cytometric assay of the cellular uptake of the polyplexes and western blot assays of the bcl2 and vegf protein expression. Transfection transit transfection reagents mirus bio. As a starting point, use 25 nm sirna final concentration in well.
We have noticed differences between these controls with regard to expression of the target gene, raising the question of the control effects on total genomic expression. These results show that xtremegene sirna transfection reagents can be used to efficiently transfect and cotransfect plasmids and sirnas into primary cells. The sirna transfection capability of the formed au denps. Xtremegene 9 dna transfection reagent is a nonliposomal multicomponent reagent for experiments involving cellular analysis. A large variety of transfection reagents have been used to deliver. However, these cells usually grow in tight clusters and are therefore more difficult to transfect than many other cell lines. Xtremegene sirna transfection reagent polymer reagent for delivering sirna to common cell lines. The x tremegene sirna transfection reagent is intended for research applications only. High efficiency versatile reagent for a wide range of common cell types most efficient versatile reagent for the widest range of cell types including difficulttotransfect cells most efficient reagent for sirna mirna delivery. Xtremegene hp dna transfection reagent sigmaaldrich.
Cd vector was evaluated by flow cytometric assay of. Enhanced delivery of therapeutic sirna into glioblastoma. For example, if the transfection reagent alters target gene expression and that is different from the transfection reagent plus the control sirna, which is the better control. Xtremegene sirna transfection reagent is a proprietary blend of lipids and other components supplied in aqueous solution, filtered through a 0. Invitrogen lipofectamine rnaimax transfection reagent 778150. Unlike for conventional dna transfection, transport of the nucleic acid into the cell nucleus is not necessary for rna transfection.
Lipofectamine rnaimax is proprietary animal originfree, chemically defined rnaispecific cationic lipid formulation designed specifically for delivery of sirna and mirna into all cell types. Transittko transfection reagent research reagent products. Dna, sirna, mirna, mrna, viral rna, crisprcas9 components, and oligonucleotides to. A lipidbased transfection reagent, such as x tremegene sirna transfection reagent, can provide a convenient, reliable, and efficient vehicle for delivering sirnas into animal cells, enabling the study of cellular and functional consequences of gene knockdown. Cells were transfected at a final concentration of 10 nm with the cy3labeled transfection. Due to its extremely low cytotoxicity, minimal need for optimization, and the ability to provide high transfection efficiency in a wide range of commonly used cell lines even in the presence of serum, it is well suited for applications in all fields of cellular analysis. For more information, please consult the appropriate material safety data sheets msdss. Out of eight different commercially available transfection reagents, there were two possible. Q i have been using the xtremegene sirna transfection reagent. Transfection reagent panel target discovery institute. Find and compare commercial and governmental sources for immunological and biological products using the linscotts directory search engine. In addition, the rnai effect caused by sirna can be detectable even after many cell divisions. The reagent keys to successful transfection experiments cationic liposomal reagents.
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