品牌 |
CELLSCRIPT | 货号 |
C-ICTAMY110510 |
产品分类 |
RNA转录试剂盒 | 研究领域 |
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产品概述 |
Store at -20°C in a freezer without a defrost cycle. Do not store at -70°C
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形式 |
The INCOGNITO™ T7 ARCA 5mC- & -RNA Transcription Kit* is optimized for high-yield synthesis of5-methylcytosine- pseudouridine-containing RNA (5mC -RNA) ARCA-capped RNA by in vitro transcription (IVT) of a DNA template that has a properly-oriented phage T7 RNA polymerase promoter usingT7-Scribe™ RNA polymerase, the canonical nucleotides ATP & GTP, the modified nucleotides 5-methylcytosine-5'-triphosphate (5mCTP) & pseudouridine-5'-triphosphate ( TP) and the anti-reverse cap analog(ARCA) m27,3'-OG[5']ppp[5']. It has been shown that 5mC -mRNAs are translated into protein at higherlevels and induce lower innate immune responses in human and other mammalian cells that expressvarious RNA sensors compared t°Corresponding unmodified mRNAs.1-4.
Because ARCA contains a 3'-O-methyl group on the m7G nucleotide (Figure 1), ARCA can only beincorporated in the correct orientation at the 5' end of the RNA during an in vitro transcription/cappingreaction.5-7 This is not true for the standard cap analog (m7G[5']ppp[5']G). Thus, ARCA incorporationresults in the synthesis of capped RNA that is more efficiently translated in vivo than standard cap analog.
Although yield varies with the DNA template and other factors, a standard 20 μl IVT reaction using 1 μg ofa control template DNA yields approximately 40 μg of 5mC -ARCA-capped RNA in 3 hours at 37°C. The5mC -ARCA-capped RNA can be enzymatically converted to a Cap 1 and 3'-poly(A)-tailed usingCELLSCRIPT's ScriptCap™ 2'-O-Methyltransferase and A-Plus™ Poly(A) Polymerase to generate5mC -mRNA for expression in cells.
The ARCA/GA 5mC PreMix contains all four ribonucleotides (GA 5mC) and the ARCA. The PreMixensures the optimal concentration of each NTP and ratio of ARCA to GTP (4:1), maximizing transcriptcapping (~80%) and yield. Because the concentration of GTP in the reaction is limiting, the ARCA ispreferentially incorporated as the first or 5'-terminal G of the transcript.
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试剂准备 |
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.
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产品描述 |
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 INCOGNITO T7 ARCA 5mC- & Ψ-RNA Transcription Kit is functionally tested under standardreaction conditions using the T7 Control Template DNA. The kit must produce at least 35 µg of RNAfrom 1 µg of the T7 Control Template DNA in 3 hours at 37°C.
Contaminating Activity Assays
All components of the INCOGNITO T7 ARCA 5mC- & Ψ-RNA Transcription Kit are free of detectableRNase and DNase activity, except for the inherent activity of the RNase-Free DNase I component.
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官网链接 |
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数量 |
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选择 | 品牌 | 货号 | 产品名称 | 规格 | 分类 | 研究领域 | 说明书 | 数量 | 目录价 | |
1 | CELLSCRIPT | C-MSC11610 | T7 mScript™ Standard mRNA Production System | 10 reactions | RNA转录试剂盒 |
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¥6600.00 | 订购 询价 | ||
2 | CELLSCRIPT | C-SCCS1710 | ScriptCap™ Cap 1 Capping System | 10 reactions | RNA转录试剂盒 |
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¥4169.00 | 订购 询价 | ||
3 | CELLSCRIPT | C-CC15011H | Cap-Clip™ Acid Pyrophosphatase | 100 units | RNA转录试剂盒 |
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¥10625.00 | 订购 询价 | ||
4 | CELLSCRIPT | C-SRI6310K | ScriptGuard™ RNase Inhibitor | 10,000 units | RNA转录试剂盒 |
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¥5865.00 | 订购 询价 | ||
5 | CELLSCRIPT | C-ASF3507 | T7-FlashScribe™ Transcription Kit | 50 reactions | RNA转录试剂盒 |
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¥6005.00 | 订购 询价 | ||
6 | CELLSCRIPT | C-ICTMY110510 | INCOGNITO™ T7 5mC- & -RNA Transcription Kit | 10 reactions | RNA转录试剂盒 |
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¥13093.00 | 订购 询价 |
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