Fugene 6 transfection reagent provides sturdy results in a wide variety of standard laboratory cell lines, making it ideal for everyday use. For use with application transfect dna into a wide variety of cell lines with high efficiency and low toxicity, for use with luciferase. Fugene 6 reagent complex prior to plating, making it a strong candidate for high throughput applications. Always mix fugene 6 reagent prior to use vortex for one second or use inversion. Improving the success of your transfection promega connections. Optimal transfection conditions for hek293 with the fugene hd reagent were determined using the protocol recommended in figure 2. Plate cells so they will be 6080% confluent at the time of transfection.
While considering a suitable transfection reagent, it is. Broad spectrum dna delivery utilize one transfection reagent and protocol for a. Citation list of established cells transfected with fugene. Although a young company, fugent has revolutionized biological study with its. Cpe wan management protocol tr069 issue 1 amendment 6. Its easytouse interface helps you to create pdf files by simply selecting the print command from any application, creating documents which can be viewed on any computer with a pdf viewer. All three offer broad spectrum transfection capability and low toxicity but the same question always comes up. An improved method for increasing the efficiency of gene transfection and transduction. This protocol is a slight modification of expression systems protocols for generating viruses. Fugene hd transfection reagent is a proprietary, nonliposomal, low toxicity formulation designed for high efficiency delivery of dna into a broad range of cell lines. Add 3 ul fugene hd directly to the transfection medium, avoid touching the side of the tube when adding fugene. I was wondering which one is better and should prefe. Teledyne lecroy is a leading provider of oscilloscopes, protocol analyzers and related test and measurement solutions that enable companies across a wide range of industries to design and test electronic devices of all types.
Download support, 10 autonomous transfer complete event. Primary culture and plasmid electroporation of the murine. The use of fugene hd does not require culture medium change or serum removal during transfection experiment. 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. Founded in madison, wisconsin in 1995 as fugent co. Teledyne lecroy analysis software protocol analyzers. Plate slightly more cells when making vsvg pseudotyped viruses. Description fugene hd transfection reagent is a proprietary, nonliposomal, low toxicity formulation designed for high efficiency delivery of dna into a broad range of cell lines.
Support for proxy protocol methods with no matching rpc 228. Selecting and isolating colonies of human induced pluripotent. In other test systems, two to threefold higher amounts of reagent yield optimal levels of expression. 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. Clinical trial protocol documents template division of aids daids for daids protocol development guidance, see the clinical trial protocol documents manual v1. Adherent cells can be trypsinized and transfected by the dna. Transfection rates were relatively high for primary cells, especially with fugene 6 20%. Pdf an improved method for increasing the efficiency of. All additional handling steps after the addition of the fugene 6 reagent. The above amounts of reagents work well with hela or cos1 cells. Use pdf download to do whatever you like with pdf files on the web and regain control.
For the reverse transfection protocol, fugene 6 and fugene hd perform as well as or better than competitor transfection reagents in terms of both transfection efficiency and cell viability. Description fugene 6 transfection reagenta is a nonliposomal reagent that transfects dna into a wide variety of cell lines with high e. Comparison of nanoparticlemediated transfection methods for. Transitlt1 transfection reagent research reagent products. The fugene hd transfection reagent protocol database has protocols for transfecting your cell line.
Follow the transfection reagent protocol for seeding cells. Fix protocol specification provides format for electronic messages and communication model fix can be used by financial institutions like brokerdealers, exchanges, institutional investors and others in the industry to communicate among each other it is widely used protocol in the financial markets industry today. Addgene is working with the trc to make this shrna cloning vector available to the scientific community. Comparable luciferase expression with the transitlt1 reagent and fugene 6 in multiple cell types. I was looking at different websites where download options are available, and always find different way to download same file. Overview of adherent cell transfection protocol for a 96well plate. Avoid contact of undiluted fugenehd transfection reagent with the sides of the tube or plate. A and b, tube 2, or fugene 6 a and b, tube 3 or nontransfection or.
A long time ago, pdf was an uncompressed file format, with the exception of images. Each reaction mix is sufficient for triplicate 96well, duplicate 24well, and single well 6 well transfections, and accounts for pipetting variations. Pdf995 makes it easy and affordable to create professionalquality documents in the popular pdf file format. The link to all protocols is a single, large pdf file that has all enls protocols in a single file. May 29, 2015 comparison of commercial transfection reagents optimized for delivery of episomal vectors into primary cells we compared the transfection efficiency and cytotoxicity of the transfection reagentepisomal vector complexes derived from the lipofectamine 2000, fugene hd and jetpei at their optimum ratios le 4, fe 4 and je 2. Finally, although reverse transfection saves time around 1 day, there is also a significant drop in transfection efficiency that must be considered. The protocol does not require removal of serum or culture medium and does not. And now, its connected to the adobe document cloud.
Oct 20, 2011 reproducibly high transfection rates with low methodologyinduced cytotoxic side effects are essential to attain the required effect on targeted cells when exogenous dna is transfected. We examined the efficiency, cytotoxicity and size of the complexes formed in the presence of different amounts of vector and dna and with variable amounts of serum. Fugene 6 transfection reagent is a proprietary blend of lipids and other components supplied in 80% ethanol, sterilefiltered, and packaged in glass vials. Free pdf to word converter has been designed to allow users to edit pdf. Its the only pdf viewer that can open and interact with all types of pdf content, including. The protocol does not require removal of serum or culture. Minimizing reagentdependent side effects as analyzed by geneexpression profiling and cytotoxicity assays article pdf available. Fugene 6 transfection reagenta is a nonliposomal reagent that transfects dna into a wide variety of cell lines with high e. Improving the success of your transfection promega. This online book describes the fundamentals of tcpip in windows vista, windows server 2008, windows xp, and windows server 2003. We offer free downloads of all our software packages.
Incubate the fugenehd transfection reagentdna mixture for 015 minutes at room temperature. Hek293 cells were grown to 85% confluency, then harvested and plated in a 96well plate at 2. Genes showing more than 5fold decrease in hdf than in human ips cells. The latest release of protocol buffers can be found on the release page. Jian yang, deshi shi, dongfang yang, xiulong song, and bingfang yan 2007 interleukin 6 alters the cellular responsiveness to clopidogrel, irinotecan and oseltamivir by suppressing the expression of carboxylesterases hce1 and hce2. Each protocol is formatted into a hyperlinked pdf file for portability and can be viewed on most devices. These protocols were developed by promega corporation or fugent, l. Fugene 6 transfection reagent is a nonliposomal reagent that transfects dna into a wide variety of cell lines with high efficiency and low toxicity. Additionally, low cell numbers can be transfected in 96well plates. The fugene hd transfection reagent is a nonliposomal formulation designed to transfect dna into a wide variety of cell lines with high efficiency and low toxicity. Promega offers three premium transfection reagents. Induction of pluripotent stem cells from adult human.
Format file system 156 minimum and maximum version. Our software packages can be used without a license key for test and evaluation. Transfection of sf9 cells in suspension nih common fund. The use of fugene hd does not require culture medium change or serum removal during. Delivery of episomal vectors into primary cells by means of. Reprogramming of human somatic cells to pluripotency with.
This policy does not apply to informed consent ic development or daids ic templates. Minimizing reagentdependent side effects as analyzed by geneexpression profiling and cytotoxicity assays. This protocol describes production of lentivirus stocks from plko shrna. Ftp is most commonly used to download files from the world wide web. For transient and stable transfection of eukaryotic cells. In animal cells, transfection is the preferred term as. Basic connectivity and graphics remoting, designed to facilitate user interaction with a remote computer system by transferring graphics display information from the remote computer to the user and transporting input from the user. Fugene 6 transfection reagent is stabilized for extended storage.
Cute pdf writer is a customizable tool that exports windowsgenerated files. Reverse transfection of human induced pluripotent stem ips cells with. Genes showing more than 5fold increase in hdf than in human ips cells. Fugene hd transfection reagent promega corporation. Transfection protocols and the reagents used to deliver nucleic acids into cells can affect cell physiology, leading to misinterpretation of results. The fugene hd protocol database is a collection of protocols that guide the transfection process for a variety of cell lines. Free excelto pdf converter allows you to easily convert your excel files to. Different approaches and modifications such as the use of nanoparticles nps are being evaluated to increase transfection efficiencies. Select the cell line, type of plate used and number of wells to be transfected and a protocol will be. Feb 01, 2006 this study compared three different synthetic reagents fugene 6, effectene and exgen 500 for the transfection of human primary myoblasts.
This is so far the most laborsaving protocol and the lentivirus yielded per 10cm dish was comparable, or even higher than several reported protocols 5,14,17. Transfection is the process of deliberately introducing naked or purified nucleic acids into eukaryotic cells. Transfection of sf9 cells in suspension december, 2008 version 1. Welcome to fugent, llc fugent, llc develops and manufactures biotechnology reagents. Promega fugene hd transfection reagent fugene hd is a novel, nonliposomal reagent for transfecting dna into a variety of cell lines with high efficiency and low toxicity. If coverslips are required, add one coverslipwell prior to adding the media. Downloads protocol buffers downloads and instructions. Adobe acrobat reader dc software is the free global standard for reliably viewing, printing, and commenting on pdf documents. Wash the cells precultured in a t25 flask with 5 ml of pbs and trypsinize with 1. Xtremegene hp dna transfection reagent sigmaaldrich. It is a protocol used to transfer files between an ftp hostserver and an ftp client computer on the internet. Wwhhen the transfer is complete, the server returns code 226 to tell the client that the transfer is complete and the data connection will be clssed.
Fugene hd transfection reagent quick protocol fb112pdf 99 kb english. A protocol for the fugene 6 transfection reagent, a nonliposomal reagent that transfects dna into a wide variety of cell lines with high efficiency and low. Feb 20, 2012 the next day substitute dmem media with media containing viral particles obtained from phoenix cells. Storage and stability fugene 6 reagent is shipped at room temperature. 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. Compared with the existing protocols, our protocol simplifies the experimental procedure and shortens the time. Fugene 6 transfection reagent protocol promega corporation.
Several studies have focused on the attained transfection efficiency. Not every application that generates pdf is equally good. Web to pdf convert any web pages to highquality pdf files. Server returns code 150 is the file status is okay and the file will be transferred. The client sends a command of retr file name in order to initiate a transfer of that file. Citation list of established cells transfected with fugene 6. Production of retroviruses using fugene 6 32100 1 on the day before transfection, plate 0. Fugene hd transfection reagenta is a novel, nonliposomal formulation designed to transfect dna into a wide variety of cell lines with high efficiency and low toxicity. Download asynchronously and put a progressbar to show the status of the download within the ui thread itself. Prepare a 6 well plate by adding 2 ml of raw cell media per well. Citation list of established cells transfected with fugene 6 transfection reagent 10t12 timothy a. This online book is a structured, introductory approach to the basic concepts and principles of the.
It may also refer to other methods and cell types, although other terms are often preferred. Figure 1 vertiga shaker manufactured by thomson instruments co. This protocol is used to generate baculoviruses containing gpcr genes of interest for expression in sf9 cells. Download tcpip fundamentals for microsoft windows from. This protocol is used to generate baculoviruses containing gpcr genes of. Olson 2000 activation of the myocyte enhancer factor2 transcription factor by calciumcalmodulindependent protein kinasestimulated binding of 1433 to histone deacetylase 5. The cells are ready for transfection after 1820 hours, or when they are about 2530% confluent. We employ researchers in the areas of organic chemistry, molecular biology, transfection, and chromatography. Add viral media 1 ml of each oct4, sox2, klf4, and cmyc media, 4 ml total supplemented with 6. Fugene 6 transfection reagent is gentle on the cells. Vwr offers transfection reagents for numerous applications, including bioproduction, in vivo work, viral production, dna and sirna transfection, crisprcas 9 and more.