Fugene 6 transfection reagent is a nonliposomal reagent that transfects dna into a wide variety of cell lines with high efficiency and low toxicity. Fugene 6 reagent into each of the first two tubes, and 6 l of the reagent into the tube labeled 6. Minimizing reagentdependent side effects as analyzed by geneexpression profiling and cytotoxicity assays article pdf available in nature methods 35 april. One the day before transfection, split 293t cells 0. While considering a suitable transfection reagent, it is. In animal cells, transfection is the preferred term as. By developing the instruments and reagents you rely on each day, we strive to support your next breakthrough so you can focus on what you do. Four distinct patterns of memory cd8 t cell responses to. The protocol does not require removal of serum or culture medium and does not require washing or changing of medium. Promega fugene 6 transfection reagent fugene 6 transfection reagent is a nonliposomal formulation designed to transfect plasmid dna into a wide variety of cell lines with high efficiency and low toxicity. Like most lipidbased reagents, it is not very good for primary and suspension cell lines. Fugene 6 get more out of your lentiviral production. Aav6 helper free packaging system cell biolabs, inc. Certain features of will be unavailable due to maintenance from saturday evening, march 21st at 8.
Fugene 6 reagent complex prior to plating, making it a strong candidate for high throughput applications. Gently shake the culture plate several times and harvest all media, including cells, in a sterile tube. Sometimes it would be better to first read the pack insert, then start working. The actual number of transfections will vary with reagent. Fugene 6 is a roche product, and appears in the roche applied science catalog. An easytouse transfection reagent that is nontoxic and gives very good results for many different cell lines. Fugene 6 transfection reagent from roche applied science. Narrow your search to look for specific document types, including application notes, manuals, protocols, and brochures. Msds fugene 6 transfection reagent, 1814443 ethanol. Virus titer was determined on lowpassagenumber tzmbl cells by. Eugene harrison roche september 22, 1928 july 28, 2004 was an american actor. Fugene 6 reagent roche molecular biochemicals as previously described 26. Fugene 6 transfection reagent is gentle on the cells.
Four distinct patterns of memory cd8 t cell responses to chronic murine cytomegalovirus infection1 michael w. Roche fugene 6 transfection reagent is a nonliposomal multicomponent reagent wellsuited for experiments involving cellular analysis. It does not contain any ingredients of human or animal origin. Search for a variety of documents via title or keyword. Cells were then additionally exposed to control medium, or various concentrations of both mse and sse for indicated periods of time. Fugene 6 transfection reagent is a nonliposomal formulation designed to transfect plasmid dna into a wide variety of cell lines with high efficiency and low toxicity. For rela and tbp tata binding protein, nuclear lysates were prepared. Vwr offers transfection reagents for numerous applications, including bioproduction, in vivo work, viral production, dna and sirna transfection, crisprcas 9 and more. Fugene 6 transfection problems transfection and transduction.
Fugene transfection protocol 1 add 3ul of fugene to 97ul of optimem, being sure to avoid touching the pipette tip to the side of the tube as the fugene will stick to plastic. After 18h of incubation, medium was replaced with high serum growth medium 30% fcs vv. The protocol does not require removal of serum or culture medium and does not require washing or changing of medium after introducing the reagentdna complex. Hill2 cmvs are herpesviruses that establish lifelong latent infection of their hosts. Mirus bio files lawsuit alleging that roches fugene 6. The complaint alleges that fugent infringes the patent, both directly and indirectly i. We celebrate the students, the researchers, the managers who work tirelessly in the pursuit of discovery. Due to the simple methodology of fugene 6 transfection reagent, minimal optimization, low cytotoxicity, and ability to provide high transfection efficiency even in the presence of serum, it is one of the standard transfection reagents in many different.
Roche fugene hd transfection reagent is designed to transfect a wide range of eukaryotic cells, including insect cells and many tumor cell lines not transfected well by other reagents e. Reverse transcriptionpcr rtpcr the first strand cdna synthesis kit invitrogen was used for reverse transcription rt. Lentiviral vectors lvv have become a valuable research tool for delivering genetic content into a wide range of cell types. Roche was born and raised in boston, massachusetts, the son of mary m. In 6cmdish, when the confluence of 293t reachs 50%,i conduct the transfection. Media and epithelia were collected and diluted appropriately and. Then i read on the rocheappliedscience homepage the following see below.
Im not sure what the relationship is between fugent and roche, nor why roche was not named in the lawsuit. Virus titer was determined on lowpassagenumber tzmbl cells by xgal staining as readout. Fugene 6 transfection reagent has been commercially available since 1997. Plate out 1x105 cells per well of a 12well plate the day before transfection. He was the original ajax man in 1970s television commercials.
Adherent cells can be trypsinized and transfected by the dna. Plate the cells such that you have 1 ml of media covering the cells. Fugene 6 transfection reagent multicomponent formulation for the transfection of eukaryotic cells cat. Since that time, its popularity has increased due to its ease of use, minimal to no cytotoxicity, and the high level of transfection in many different cell lines. One 1ml vial of fugene hd transfection reagent contains sufficient material to transfect over 3,000 wells in 96well cell culture plates at a 3. Viruses were harvested at 48h and 72h posttransfection. Additionally, low cell numbers can be transfected in 96well plates. It may also refer to other methods and cell types, although other terms are often preferred.
Storage and stability fugene 6 reagent is shipped at room temperature. Acebron, emil karaulanov, carmen reinhard, dierk ingelfinger, michael boutros, christof niehrs to whom correspondence should be addressed. Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Sigma aldrich, using fugene 6 roche, berlin, germany in optimem medium gibco. Cas 64175 fugene 6 transfection reagent, 1814443 ethanol msds toxicity property. Transfect 293t cells with plasmids using fugene 6 1. Fugene 6 transfection reagent university of pittsburgh. 5 m edta to a final of 10 mm to the plate and incubate for 3.
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