MessageMAX™ T7 ARCA-Capped Message | C-MMA60710 | CELLSCRIPT | Leading Biology Inc.

来自 : 发布时间：2025-01-30

AbcamAbD SerotecActive MotifAtlas AntibodiesAdipogenAgriseraAllele biotechnologyAbnovaAllerMAbsAnogenAbbexaAbgentAbbkineAbseaACROBiosystemsAdvanstaAffinity BiosciencesAlomoneAmsbioAnaSpecAssayProApexBioABL (Advanced BioScience Laboratories)AssayBioTechArthus BiosystemsAgrenvecAdvanced Targeting SystemsAccelagenAbsolute antibodyAxolbioAlpha Diagnoestic InternationalAAT BioquestAmerican Research ProductsARBOR ASSAYSAvidityAbbiotecAbcoreAccuStandardAdvanced BioMatrixAthens ResearchAnatraceAvanti Polar LipidsATCCAG ScientificArcticzymesAstartebioAllele BiotechAvantamaAves LabAviscera BioscienceApollo ScientificApplied BiophysicsAnnoron The MessageMAX™ T7 ARCA-Capped Message Transcription Kit produces capped RNA by in vitrotranscription using T7 RNA polymerase and the anti-reverse cap analog (ARCA) m27,3 -OG[5 ]ppp[5 ]. A30 minute, 20 μl MessageMAX reaction yields up to 60 μg of ARCA-capped RNA from 1 μg of the 1.4 kbstandard T7 control template DNA. Because ARCA contains a 3 -O-methyl group on the m7G nucleotide(Figure 1), ARCA can only be incorporated in the correct orientation at the 5 end of the RNA during anin vitro transcription/capping reaction.1-3 This is not true for the standard cap analog (m7G[5 ]ppp[5 ]G).Thus, ARCA incorporation results in the synthesis of capped RNA that is more efficiently translatedin vivo than standard cap analog.The ARCA/NTP PreMix contains all four ribonucleotides and the ARCA. The PreMix ensures the optimalconcentration of each NTP and ratio of ARCA to GTP (4:1), maximizing transcript capping (~80%) andyield. Because the concentration of GTP in the reaction is limiting, the ARCA is preferentiallyincorporated as the first or 5 -terminal G of the transcript.CELLSCRIPT also offers the INCOGNITO™ T7 ARCA 5mC- -RNA Transcription Kit for theproduction of 5-methylcytosine- pseudouridine-containing (GA m5C) ARCA-capped RNA. Such RNAhas been used in the landmark publication by Warren et al. (2010), to successfully reprogram multiplehuman cell types into induced pluripotent stem cells.4 It has been shown that -mRNAs and 5mCmRNAs are translated into protein at higher levels and induce lower innate immune responses in humanand other mammalian cells that express various RNA sensors compared t°Corresponding unmodifiedmRNAs. RNase-Free DNase I is provided in a 50% glycerol solution containing 50 mM Tris-HCl, pH 7.5,10 mM CaCl2, 10 mM MgCl2 and 0.1% Triton® X-100. ScriptGuard RNase Inhibitor is supplied in a50% glycerol solution containing 50 mM Tris-HCl, pH 7.5, 100 mM NaCl, 10 mM DTT, 0.1 mM EDTAand 0.1% Triton® X-100. All other enzymes are provided in a 50% glycerol solution containing 50 mMTris-HCl, pH 7.5, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA and 0.1% Triton X-100. Unit Definitions One unit of RNase-Free DNase I digests 1 μg of pUC19 DNA to oligodeoxynucleotides in 10 minutesat 37°C.One unit of ScriptGuard RNase Inhibitor results in 50% inhibition of 5 ng of RNase A. Activity ismeasured by the inhibition of hydrolysis of cyclic 2 ,3 -CMP by RNase A.Functional Testing The MessageMAX T7 ARCA-Capped Message Transcription Kit is functionally tested under standardreaction conditions using the T7 Control Template DNA. The kit must produce at least 40 μg of RNAfrom 1 μg of the T7 Control Template DNA in 30 minutes at 37°C.Contaminating Activity Assays All components of the MessageMAX T7 ARCA-Capped Message Transcription Kit are free ofdetectable RNase and DNase activity, except for the inherent activity of the RNase-Free DNase Icomponent. 安诺伦官方微信

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