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Invitrogen™ mMESSAGE mMACHINE™ T7 mRNA Kit with CleanCap™ Reagent AG
Description
The mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG contains all the reagents and buffers necessary for high quality in vitro synthesis of large amounts of mRNA with the naturally occurring Cap 1 structure. Utilization of the CleanCap Reagent AG ensures superior results relative to older capping technologies and greater protein production. Each standard 20-μL reaction yields a minimum of 100 μg of Cap 1-containing mRNA, while the scaled-up protocol yields up to 1 mg of mRNA. This kit is suitable for full- or partially-modified-nucleotide substitution to generate modified mRNA.
Features of the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG include:
- Contains proven CleanCap capping technology
- Optimized to achieve high mRNA yields (>5 mg/mL)
- Produces mRNA that is over 95% capped
- Optimized for production of mRNA from 0.9–10 kb templates
- Minimum yields of 100 μg per reaction, up to 150 μg per reaction (see figure below)
- Allows use of modified nucleotides, full- or partial-modified substitution
- Includes linearized 0.9 kb GFP control template, yields ≥100 μg mRNA
- Made up of high quality TheraPure reagents to enable transition from RUO to GMP
- Available in two sizes: 50 and 1000 reactions*
*Note: 1000 reaction kit is designed to support scale-up to large amounts of mRNA production enabling ≥100 mg mRNA.
The mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG contains the trinucleotide cap analog CleanCap Reagent AG, which has the Cap 1 structure and can be used for co-transcriptional capping of mRNA without compromising yield. This reagent produces high capping efficiencies compared to legacy capping reagents such as ARCA. Cap 1-containing mRNAs are considerably more stable and have superior translational activities compared to Cap 0 mRNAs produced using ARCA or mCap cap analogs.
A linearized DNA template containing an AG-initiating T7 RNA polymerase promoter is strongly recommended for this kit to achieve mRNA yields >5 mg/mL with over 90% of the transcripts capped. The mRNA produced with this kit can be used for many applications, including transfections, preclinical mRNA therapeutic mRNA studies, preclinical vaccine development, microinjections, in vitro translation, and RNA structure and function analysis.
Specifications
Specifications
| Cell Type | HEK293, 293, HeLa, CHO-K1, CHO, Jurkat, Primary Human T Cells, Primary Cells, Stem Cells, iPSC, NIH-3T3, HepG2, A549, HUVEC |
| Final Product Type | Synthetic mRNA, mRNA |
| Format | Animal Origin Free Kit |
| Isolation Technology | Silica Spin Column |
| Purification Target | mRNA, Synthetic mRNA |
| Sample Type | mRNA, Synthetic mRNA |
| No. of Reactions | 50 Reactions |
| Product Line | mMESSAGE mMACHINE |
| Product Type | T7 In Vitro Transcription kit |
| Quantity | 50 Reactions |
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Frequently Asked Questions (FAQs)
CleanCap is a new capping technology that can provide higher mRNA yields and capping efficiencies than ARCA. CleanCap contains the cap 1 structure, which is found in higher eukaryotes such as mice or humans. ARCA contains the cap 0 structure that is found in lower eukaryotes such as yeast. Cap 1 mRNA can have superior translational activities in humans compared to Cap 0 mRNAs.
The RNA can be introduced into cells by electroporation or transfection. Our Neon NxT Electroporation System can be used to introduce the RNA by electroporation. Our Lipofectamine MessengerMAX Transfection Reagent is designed for mRNA delivery and works with many cell types.
The RNA synthesized using the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621) can be purified using the provided LiCl Precipitation Solution. We recommend referring to the user guide for details. Alternatively, the RNA can be purified using spin columns such with the MEGAclear Transcription Clean-Up Kit (Cat. No. AM1908). Note that the MEGAclear Transcription Clean-Up Kit can purify up to 500 µg RNA and that it only purifies RNAs >100 nucleotides. If shorter RNAs need to be purified, we recommend using the GeneJET RNA Cleanup and Concentration Micro Kit (Cat. No. K0841).
Yes, modified nucleotides can be used. We recommend referring to the user guide for guidance on this.
Yes, we recommend using a template encoded poly-A tail as it yields better expression than when a poly-A tail is added enzymatically using poly-A polymerase. Refer to the user guide for guidance on making a template with a poly-A tail.
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