Invitrogen™ mMESSAGE mMACHINE™ T7 Transcription Kit

Product Code. 10438644

Brand: Invitrogen™ AM1344

Description

mMESSAGE mMACHINE kits are designed for the in vitro synthesis of large amounts of capped RNA. Capped RNA mimics most eukaryotic mRNAs found in vivo, because it has a 7-methyl guanosine cap structure at the 5' end.

mMESSAGE mMACHINE kits are designed for the in vitro synthesis of large amounts of capped RNA. Capped RNA mimics most eukaryotic mRNAs found in vivo, because it has a 7-methyl guanosine cap structure at the 5' end. mMESSAGE mMACHINE kit reactions include cap analog [m7G(5')ppp(5')G] in an ultra high-yield transcription reaction. The cap analog is incorporated only as the first or 5' terminal G of the transcript because its structure precludes its incorporation at any other position in the RNA molecule.

How mMESSAGE mMACHINE kits work

mMESSAGE mMACHINE kits have a simplified reaction format in which all four ribonucleotides and cap analog are mixed in a single solution. The cap analog:GTP ratio of this solution is 4:1, which is optimal for maximizing both RNA yield and the proportion of capped transcripts. mMESSAGE mMACHINE kits are ideal for the routine synthesis of capped RNAs for oocyte microinjection, in vitro translation, transfection, and other applications. The high yields are achieved by optimizing reaction conditions for RNA synthesis in the presence of high nucleotide concentrations. In addition, the RNA synthesized is protected from degradation by any contaminating ribonucleases that may be present with RNase inhibitor—a component of the Enzyme Mix.

Using mMESSAGE mMACHINE Kits

In a 20 μL reaction during a two hour incubation, mMESSAGE mMACHINE high yield capped RNA transcription will yield large mass amounts of capped RNA. Up to 10–50 times the yield obtained with conventional in vitro transcription reactions. The ratio of cap analog to GTP has been optimized to allow the best compromise between yield (15–35 μg) and proportion of transcripts that are capped (80%).

mMESSAGE mMACHINE kits also contain a LiCl precipitation solution that is efficient for separating proteins and unincorporated nucleotides (including free cap analog) from the capped RNA, allowing an increased efficiency of translation. mMESSAGE mMACHINE kits are only optimized for use with the polymerases included in the kit. Using a different polymerase may result in low yields. The mMESSAGE mMACHINE kits contain all the buffers and reagents necessary for 25 transcription reactions. Using the control template supplied with the kits (Xenopus elongation factor 1α, pTRI Xef), each mMESSAGE mMACHINE reaction will yield approximately 20–30 μg of RNA using T3 or T7 RNA polymerase or about 15–25 μg RNA using SP6 RNA polymerase.

Specifications

• Content And Storage: T7 Enzyme Mix, 10X Reaction Buffer 2XNTP/CAP Solution, GTP Solution, pTRI-Xef, TURBO DNase, Ammonium Acetate Stop Solution, Lithium Chloride Precipitation Solution, and Gel Loading Buffer II are all stored at –20°C. Nuclease-free Water may be stored at any temperature.
• Final Product Type: Synthetic mRNA, mRNA
• Format: Kit
• Sample Type: mRNA, Synthetic mRNA
• For Use With (Application): In Vitro Transcription, synthetic mRNA, mRNA, transfection, LNPs, GENEart, linearized DNA plasmid, delivery, PCR products
• No. of Reactions: 25 Reactions
• Product Line: Ambion™, mMESSAGE mMACHINE™
• Product Type: T7 In Vitro Transcription kit
• Quantity: 25 Reactions
• Promoter: T7
• Shipping Condition: Dry Ice
• Yield: 20-30 μg
• Starting Material: PCR Products, Linearized DNA plasmid

Safety and Handling

Product Identifier

• mMESSAGE mMACHINE T7 Transcription Kit

Signal Word

• Danger

Hazard Category

• Acute toxicity Category 4
• Carcinogenicity Category 2
• Serious eye damage/eye irritation Category 2
• Reproductive toxicity Category 1B
• Skin corrosion/irritation Category 2
• Specific target organ toxicity after repeated exposure Category 2

Hazard Statement

• H302-Harmful if swallowed.
• H315-Causes skin irritation.
• H319-Causes serious eye irritation.
• H351-Suspected of causing cancer.
• H360-May damage fertility or the unborn child.
• H373-May cause damage to organs.

Precautionary Statement

• P201-Obtain special instructions before use.
• P202-Do not handle until all safety precautions have been read and understood.
• P260-Do not breathe dust/fume/gas/mist/vapours/spray.
• P264-Wash hands thoroughly after handling.
• P270-Do not eat, drink or smoke when using this product.
• P280-Wear protective gloves/protective clothing/eye protection/face protection.
• P301+P310-IF SWALLOWED: Immediately call a POISON CENTER/doctor /
• P302+P352-IF ON SKIN: Wash with plenty of water/soap.
• P305+P351+P338-IF IN EYES: Rinse cautiously with water for several minutes. Remove contact lenses, if present and easy to do. Continue rinsing.
• P308+P313-IF exposed or concerned: Get medical advice/attention.
• P314-Get medical advice/attention if you feel unwell.
• P330-Rinse mouth.
• P332+P313-If skin irritation occurs: Get medical advice/attention.
• P501b-Dispose of contents/container in accordance with local/regional/national/international regulations.

Supplemental information

• MIXTURE LIST-Contains : lithium chloride, formamide

For Research Use Only. Not for use in diagnostic procedures.