Does thermo fisher scientific offer a proteasefree dnase. Ambion turbo dnafree dnase treatment and removal reagents are designed to remove. Thermo scientific dnase i, rnasefree is an endonuclease that digests single and doublestranded dna. Western blotting using the invitrogen nupage novex bis. Centrifugation, polymerase chain reaction, precipitation. Dnase i is suitable for removing dna from protein preparations, nick translating dna, and generating random fragments for dideoxy sequencing. If using dnase i, hc, enzyme can be diluted in 1x dnase reaction buffer just prior to use, or in storage buffer not supplied see composition on reverse page for longer storage. Pdf dnase concentration assay to obtain dnafree rna from. Electroporation using neon electroporation from invitrogen. Dnase i deoxyribonuclease i digests single and doublestranded dna to. If two or more fail to knock down your target rna by at least 70% under these conditions, invitrogen will design and ship a fourth stealth rnai sirna to your target for free. Dnase i amplification grade from invitrogen,dnase i amplification grade digests single and doublestranded dna to oligodeoxyribonucleotides. Thermo scientific dnase i, rnase free is an endonuclease that digests single and doublestranded dna.
The phosphopeptide positive control zlyte tyr 4 phosphopeptide and the nonphosphorylated negative. Dnase i recombinant, rnasefree from bovine pancreas. The enzyme also has a 6fold lower k m for dna, thus enabling effective removal of trace quantities of dna contamination. Fill plastic neon tube with 3 ml of invitrogen electrolyte buffer and insert tube into the neon pipette station. Real time pcr for gene expression analysis isolate rna using hot acid phenol protocol. The ambion dnafree dnase treatment and removal reagents are designed to remove contaminating. Gibco cell dissociation buffer enzyme free pbsbased from invitrogen,cell dissociation buffers are membranefiltered isotonic and enzymefree aqueous formulations of salts chelating agents and cell. Service see page 66 or download the manuals from our web site at. Dna from the sources listed has been successfully isolated and used to produce pcr products or for southern blot experiments. Studies of dnaprotein interactions by dnase i, rnase free footprinting 1. Advanced granulation technology agt media are a dry form of cell culture media in a granular format suited to industrialscale production applications which invitrogen can now make in an animalorigin. To work with larger amounts of rna, scale up the reaction including volume linearly.
Turbo dnafree kit turbo dnase treatment and removal. Dnase i functions by hydrolyzing phosphodiester linkages, producing mono and oligonucleotides with 5phosphate and 3hydroxyl group. Service see page 51 or you may download the manuals from our web site at. Dnase i amplification grade from invitrogen, dnase i amplification grade digests single and doublestranded dna to oligodeoxyribonucleotides. Volumes of the reaction mixture and 50 mm edta solution. Can someone provide a protocol for dnase i treatment after.
Dnase provided in the kit is overexpressed in an animal free system, and is then extensively. Select the desired running buffer mops works for 200 to 14 kda and mes for 60 to 2. Invitrogen brings animalorigin free media formulation to new. The blockit rnai designer is such an effective tool for the design of stealth rnai sirna that if you order the three best stealth rnai sirna sequences designed by the blockit rnai designer, we guarantee that two of them will give greater than 70% knockdown of mrna, given that the transfection efficiency in your experiment is at least 80%. Our balanced salts include dulbeccos phosphate buffered saline dpbs. After about 3 years of free software distribution you have to pay for the license.
Advanced granulation technology agt media are a dry form of cell culture media in a granular format suited to industrialscale production applications which invitrogen can now make in an animalorigin free environment in response to fears such as prion and microbe contamination. Dnase i amplification grade is specifically purified and tested for absence of rnase contamination for superior rtpcr 1. Dnase i amplification grade from invitrogen biomedicine. Removal of template dna following in vitro transcription 1, see protocol on reverse page. The turbo dnafree kit contains reagents for the efficient, complete.
It is also available as a turbo dnafree kit, which utilizes an ambionengineered hyperactive. Pdf the success of gene expression studies, protein synthesis, and construction of cdna libraries directly. Treated and untreated samples were reverse transcribed with life technologies. Endonuclease g modulates the alternative splicing of. Dnase concentration assay to obtain dnafree rna from sugarcane leaves. Earles and hanks balanced salt solutions ebss hbss. For optimization of sirna delivery, the following silentmer validated. Dnafree dnase treatment and removal reagents are designed for the removal of contaminating dna from rna samples and for the removal of dnase after. Simple and rapid optimization with maximum transfection. Easy dna kit the easy dna kit is a simple, quick, and inexpensive method for the isolation of dna from a variety of sources. Inactivate the dnase i by the addition of 1 l of 25 mm edta solution to the reaction mixture. Invitrogen catalog first page, datasheet, datasheet search, data sheet, datasheets, datasheet search site for electronic components and semiconductors, integrated circuits, diodes, triacs, semiconductors.
Generation of a library of randomly overlapping dna inserts. We get good results with the cdna synthesis after weve used this dnase. The gels have a shelf life of 48 weeks depending upon. Invitrogen corporation invitrogen bv 1600 faraday avenue po box 2312, 9704 ch. Learn more about how this product is being used in the product citation tool. This is a free resource for the scientific community that is compiled by addgene. Invitrogen ambion dnase i rnasefree 2000 units products. Jan 11, 2010 the dnafree kit from ambion comes in the size of 50 rxns, complete with rnasefree dnase i, an optimized 10x reaction buffer, and a novel dnase removal reagent, with or without a user manual, which can also be downloaded from an online source. Hyperactive turbo dnase is a catalytically superior enzyme compared to wild type. In addition, these proteins can also associate with claudin, occludin and factin, at tight junction stands, where they provide a linkage between the actin cytoskeleton and the tight junction. Protocol for dnase i treatment of rna molecular biology. This invitrogen novex powerease 500 power supply is used and in excellent condition.
Ambion turbo dnafree dnase treatment and removal reagents are designed. Plate cells so they will be 7090% confluent at the time of transfection. Dna labeling by nicktranslation in conjunction with dna polymerase i 1, see protocol on reverse page. Flow cytometry results within 72 h for cells transfected with a pendoggfp, c. Dnase i from bovine pancreas is a glycoprotein of mr 37000. Invitrogen corporation invitrogen bv 1600 faraday avenue po box 2312, 9704 ch groningen carlsbad, ca 92008 the netherlands toll free tel. Invitrogen brings animalorigin free media formulation to. Name vector type resistance marker bacterial resistance source sequence available. For the optimization of plasmid delivery, a plasmid dna expressing the luciferase gene pcmviluc was. A special procedure is used to remove rnases from the dnase preparation.
The kit also includes an optimized dnase reaction buffer. Applicationsdnase i is suitable for removing dna from protein preparations, nick translating dna, and generating random. That one has an edta deactivation step, using 1ul of 25mm edta for each 10ul sample. The assay was carried out according to the kit instructions. Invitrogen qubit 4 manuals manuals and user guides for invitrogen qubit 4. Invitrogen novex powerease 500 in stock, we buy sell. You will need reactions for each sample with each primer set. Inhibition of nuclease activity by a spliceswitching. Turbo dna free kit is similar to the dna free kit but includes turbo dnase, an engineered hyperactive dnase that exhibits up to 350% greater catalytic efficiency than wild type dnase.
Place electrode wick within the black alignment marks at both ends of the cassette. Dnafree kit dnase treatment and removal reagents user. Dnase i, amplification grade thermo fisher scientific. A, d levels of fulllength ndnase i splice variant, b, e. Seal all sample wells both sides with purple invitrogen sticker. Invitrogen creates products for cellular, protein and molecular biology research that allow scientists to innovate. This unit was made by novex which was incorporated into invitrogen. Rna was isolated with the concert plant rna reagent invitrogen, thermo fisher. Preparation of dnafree rna prior to rtpcr and rtqpcr 2, see protocol on reverse page.
It functions by hydrolyzing phosphodiester linkages, producing mono and oligonucleotides with a 5phosphate and a 3hydroxyl group. The dnafree kit from ambion comes in the size of 50 rxns, complete with rnasefree dnase i, an optimized 10x reaction buffer, and a novel dnase removal reagent, with or without a. Ribonuclease has been reduced to nondetectable levels. Details of the multiple cloning sites for pcdna z 3. Flow cytometry results within 72 h for cells transfected with a. The ambion dnafree dnase treatment and removal reagents are designed to.
Dnase i is a dnaspecific endonuclease that hydrolyzes ds or ssdna. Dnase i, amplification grade, digests single and doublestranded dna to oligodexyribonuleotides containing a 5 phosphate. In the invitrogen nupage bistris discontinous buffer system, the electrophoretic mobility of the proteins and the subsequent separation range of the gel is dependent on two factors. Welcome to vector database vector database is a digital collection of vector backbones assembled from publications and commercially available sources. For the optimization of plasmid delivery, a plasmid dna expressing the luciferase gene pcmviluc was used. Dnase i, amplification grade, is suitable for eliminating dna during critical. We have 1 invitrogen qubit 4 manual available for free pdf download. Dnase i, amplification grade, is suitable for eliminating dna during critical rna purification procedures such as those prior to rnapcr amplification. Incubation of 2 units of dnase i with 10 g of singlestranded rna ladder for 1 hour at. Rnase free dnase i is recommended to degrade dna in presence of rna, in absence of rnase is critical to maintain integrity of rna.
Incubation of 100 units of dnase rnase free i with 10 g of doublestranded rna ladder for 2 hours at 37c resulted in the same electrophoretic profile as untreated rna. Dna from the sources listed has been successfully isolated and used to. Our classical media include original formulae and variations of bme, dulbeccos modified eagles medium dmem. Invitrogen instructions say to use buffer e if electroporating in 10 l and e2 if electroporating in 100 l. Gibco cell dissociation buffer enzyme free pbsbased from. This protocol is designed to remove trace to moderate amounts of contaminating dna. It hydrolyzes phosphodiester bonds producing mono and oligodeoxyribonucleotides with 5phosphate and 3oh groups.
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