Tpro pfect transfection reagent is a broad spectrum, low toxicity transfection reagent that provides high efficiency plasmid dna delivery in many mammalian cell types. Plus reagent is a proprietary reagent for precomplexing dna that enhances cationic lipidmediated transfection of dna into many cultured eukaryotic cells. During operation, always work in a fume hood, always wear a lab coat, disposable gloves, protective goggles and antifog procedure mask. Tpro pfect transfection reagent product information. Invitrogen trizol reagent is a readytouse reagent, designed to isolate high quality total rna as well as dna and proteins from cell and tissue samples of human, animal, plant, yeast, or bacterial origin, within one hour. Trizol reagent is a complete, readytouse reagent for the isolation of highquality total rna or the simultaneous isolation of rna, dna, and protein from a variety of biological samples. The invitrogen life technologies trizol reagent total rna isolation reagent is a readytouse reagent for the isolation of total rna from cells and tissues for use in pcr analysis. Oragenerna purification protocol using tri reagent ls trizol. Ribozol rna extraction reagent contains phenol, which is a poison and can cause burns.
Life technologies is now part of thermo fisher scientific. Lipofectamine 3000 reagent maintains a high transfection efficiency within a robust dynamic range of lipid doses for quick and easy optimization. However, i go the same values for free glucose and. Trizol ls reagent from invitrogen, trizol reagents are readytouse monophasic solutions of phenol and guanidine isothiocyanate suitable for isolating total rna dna and proteins 16. Use a chemical fume hood in order to avoid breathing the vapor. This protocol is to produce lentivirus stocks from hairpinplko. Sep 18, 2007 the protocol is easy to follow and allows interruption after the homogenization when samples can be frozen at 80c, if necessary. Trizol reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components which. Total rna isolated by trizol reagent is free of protein and dna contamination. Nuclease free water 8 mm naoh 1% sds protocolprocedure.
Technical bulletin product description tri reagent is a quick and convenient reagent for use in the simultaneous isolation of rna, dna, and protein. Simplified instructions for invitrogen nupage gels running nupage gels 1. The reagent is an improvement of the singlestep rna isolation method developed by chomczynski and sacchi. Tpro pfect is a low toxicity, serumcompatible transfection reagent that eliminates the need for. Trizol solubilization and extraction is a relatively recently developed general method for deproteinizing rna. Altogen cro offers in vivo rnai services, tumor xenograft models, toxicology testing, stable cell line generation, and. Please use the netaffx analysis center to limit download data to your probesets of interest. The pureyield rna midiprep systema is a columnbased method for the isolation of essentially dna free total rna directly from biological samples 1. Do not add qiagen proteinase k directly to buffer al. Briefly, cryopreserved cells were thawed and gently pelleted. The reagent, a monophasic solution of phenol and guanidine isothiocyanate, is an improvement to the singlestep rna isolation method developed by chomczynski and sacchi 1. Trizol is simple to use and suitable for the isolation of rna from small to large quantities of tissue 50100 mg to greater than 1 mg and cells 5 x 10 6 to greater than 10 7, of human, animal, plant or bacterial origin.
Optimem serum free media life technologies, invitrogen. Tri reagent for processing tissues, cells cultured in monolayer or cell pellets catalog number t9424 store atroom temperature. The kit utilizes the strong lysis capability of tripure reagent. Homogenize 10 to 50 mg of frozen muscle piece in 1ml of tri reagent in falcon tube. Dnareagent complexwell 10 l 50 l 250 l incubate cells for days at 37c amount 96well 24well 6well dnawell 100 ng 500 ng 2500 ng lipofectamine 2000 reagent well 0. Invitrogen trizol plus rna purification kit provides a simple, reliable, and rapid method for isolating highquality total rna from a wide variety of samples, including animal and plant cells and tissue, bacteria, and yeast.
Trizol reagent can also be used with phasemaker tubes to isolate rna. Description of protocols two protocols are provided in this handbook. For emergencies concerning 7335 executive way human health or the environment. Genezol reagent is a phenol, chloroform and guanidine isothiocyanate based scalable solution for extracting highquality total rna as well as simultaneous extraction of rna, dna and protein from a wide variety of samples such as blood, buffy coat, plasma, serum, cultured cells and tissue. This method is particularly advantageous in situations where cells or tissues are enriched for endogenous rnases or when separation of cytoplasmic rna from nuclear rna is impractical. Use of the transfect protocol database is free of charge and no registration is required. Rna was extracted from tumor samples using trizol invitrogen, carlsbad, ca according to the manufacturers instructions invitrogen. Trizol reagent allows the user to perform sequential precipitation of rna, dna, and proteins from a single sample chomczynski, 1993. Trizol ls reagent is a monophasic solution of phenol, guanidine isothiocyanate, and other proprietary components. The labserv range of nitrile powderfree gloves have been specifically designed for. Transfection experiments require standard cell culture reagents and. The extracted rna can be used directly in a variety of downstream applications such as cdna library.
Tripure reagent maintains the integrity of the rna, while disrupting cells and dissolving cell components. The supernatant was removed and the pellet was dispersed while adding trizol with vortexing 1 ml trizol1x107 cells. Chemical product and company information life technologies, inc. Genezol reagent contains phenol and guanidine isothiocyanate. Invitrogen trizol reagent 100ml invitrogen trizol reagent. Successful isolations from human, animal, plant, yeast. The supernatant was removed and the pellet was dispersed while adding trizol with vortexing 1 ml trizol 1x107 cells. Tri reagent solution also sold as trizol is a mixture of a mixture of guanidine thiocyanate and phenol in a monophase solution that is used for the isolation of dna, rna and protein from biological samples of human, animal, plant, yeast, bacteria, and virus.
Trizol plus rna purification kit thermo fisher scientific. The equivalent of 1ml of regular trizol is 250ul of h 2 o and 750ul of trizol ls. Disposablenondisposable glassware, plasticware and automatic pipettes should be sterile rnase free and used only for rna procedures. Invitrogen lipofectamine 3000 transfection reagent fisher.
Oragenerna purification protocol using tri reagent ls trizol introduction tri reagentls is a quick and convenient reagent for use in the isolation of rna from liquid samples of human origin. Transfection reagents are highly efficient for dna and sirna transfection in vivo and in vitro. This protocol is optimized for use with 100 ngwell plasmid and 100. Efficiently transfect difficulttotransfect cells lipofectamine 3000 reagent is designed to efficiently transfect difficulttotransfect cells, yielding superior transfection performance across the broadest array of. Triazole reagent msds search our sds online database free.
Service see page 66 or download the manuals from our web site at. Trizol reagent is a monophasic solution of phenol and guanidine isothiocyanate. Purification of rna using trizol tri reagent request pdf. Isolation of total rna with trizol reagentlife technologies. The procedure is an adaptation of the single step method reported by chomczynski and sacchi for total rna isolation. Use plus reagent to enhance transfection results with lipofectamine ltx reagent, lipofectamine reagent, lipofectin reagent, cellfectin reagent, and oligofectamine reagent. For rna and protein isolation, refer to trizol reagent user guide pub. Keep all the tubes cold till homogenization is over. Leave the tubes at room temperature for 5 minutes to permit complete dissociation of nucleoprotein complexes.
Your new online msds binder is a place for you to store the material safety data sheets you need to deploy. Invitrogen trizol reagent is a readytouse reagent, designed to isolate high. Analysis of mouse liver glycogen content bioprotocol. Invitrogen trizol ls reagent is a readytouse reagent, designed to isolate high quality total rna as well as dna and proteins from cell and tissue samples of human, animal, plant, yeast, or bacterial origin, within one hour. Trizol reagent invitrogen, carlsbad, ca usa was used for extraction of total rna from the solutions, adopting the methodology described by the manufacturer rio et al. The folinciocalteu reagent fcr or folins phenol reagent or folindenis reagent, also called the gallic acid equivalence method gae, is a mixture of phosphomolybdate and phosphotungstate used for the colorimetric in vitro assay of phenolic and polyphenolic antioxidants. Please do not make copies of or distribute this protocol. Depctreated water ambion trizol reagent invitrogen ice cold pbs cell scraper 70% ethanol isopropyl alcohol b. The procedure is an adaptation of the single step method reported by. All tissue and culture samples were extracted using a previously described guanidiniumisothyocyanatechloroform trizol reagent, invitrogen, carlsbad, ca, usa protocol 51. Trizol ls reagent from invitrogen,trizol reagents are readytouse monophasic solutions of phenol and guanidine isothiocyanate suitable for isolating total rna dna and proteins 16. Refer to trizol reagent and phasemaker tubes complete system user guide pub. Product catalogue thermo fisher scientific new zealand. Tetramethylrhodamine tmrm thermo fisher scientific.
Other companies are charging thousands of dollars to set up accounts and give you access to their msds online database. The rna precipitate, often invisible before centrifugation, forms a gellike pellet on the side and bottom of the tube. This monophasic solution of phenol and guanidine isothiocyanate is designed to isolate separate fractions of rna, dna, and proteins from cell and tissue. Invitrogens website contains the wellexplained protocol with hints for working with trizol and rna and faqs about the method.
Extraction and purification of total rna using trizol or. May 12, 2009 the reagent is an improvement of the singlestep rna isolation method developed by chomczynski and sacchi. Polymer in vivo transfection reagent altogen biosystems. Sacchi developed a singlestep method of rna isolation by acid guanidinium thiocyanatephenolchloroform extraction. Homogenize the samples using a sterile or rnase free plastic, glass pestles or power. Maximizing total rna yield from trizol reagent protocol. Versatileone reagent for dna, rna, and cotransfection. The kit utilizes the strong lysis capability of trizol reagent, followed by a convenient and timesaving silica. Affymetrix support by product for genechip human transcriptome. I have tried the acidhydrolysis method to quantify glycogen in the liver of mice following your protocol. Browse our database of protocols for cell culture, nucleic acid, protein analysis, rna and dna applications, cloning, cell analysis, and drug discovery research.
Refrigerated centrifuge microcentrifuge micropipettors aerosolbarrier tips vortex mixer powder free gloves centrifuge tubes. Purification of dna from saliva stabilized in rnaprotect. Rna isolation with trizol invitrogen and qiagen rnaeasy. Tetramethylrhodamine, methyl ester tmrm is a cellpermeant dye that. Trizol or trizol ls reagent from invitrogen trizol ls is specifically created to work with liquid samples i. Extraction and purification of total rna using trizol or tri. Add 1 ml of trizol or tri reagent to every 50100 mg of tissue. Select the desired running buffer mops works for 200 to 14 kda and mes for 60 to 2.
Purification of dna from saliva stabilized in rnaprotect saliva reagent this protocol describes how to purify dna from human saliva samples stabilized in rnaprotect saliva reagent the reagent provides immediate stabilizati on of both dna and rna at room temperature. Isolation of rna from tissue and cells is an essential tool to study gene expression. Invitrogen lipofectamine 3000 transfection reagent. For preparation of standard curve, glucose solutions of 0. Trizol reagent replaces 82498 life technologies, inc. Lipofectamine 2000 dna transfection reagent protocol.
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