Invitrogen™ mMESSAGE mMACHINE™ T7 ULTRA Transcription Kit

Description

The mMESSAGE mMACHINE T7 ULTRA Transcription Kit generates RNA transcripts with higher protein yields due to the combination of the anti-reverse cap analog (ARCA) with patented transcription reagents.

The mMESSAGE mMACHINE T7 Ultra Kit combines the anti-reverse cap analog (ARCA) with a patented high-yield transcription technology to generate RNA transcripts that produce higher protein yields compared to other transcripts upon translation.

Features of the mMESSAGE mMACHINE T7 Ultra Kit include:

• Synthesize more protein from in vitro-transcribed RNA
• Express RNA transcripts more efficiently both in vitro and in vivo
• Stabilize transcripts in vivo with included reagents for poly(A) tailing

ARCA-capped RNA is translationally more active

A base modification in ARCA results in its incorporation in the functional, translatable orientation only; traditional cap analog can be incorporated in both functional and nonfunctional orientations. As a result, incorporating ARCA into transcription reactions yields capped RNAs that are 100% translatable. This is further enhanced by the inclusion of poly(A) tails in mMESSAGE mMACHINE T7 Ultra transcripts. Experiments comparing ARCA and ARCA/poly(A)-tailed transcripts to cap analog and cap analog/poly(A)-tailed transcripts indicate higher levels of protein synthesis with ARCA capped RNA.

What Is ARCA?

ARCA is a modified cap analog in which the 3' OH group (closer to m7G) is replaced with OCH₃ (see schematic). Because of this substitution, the RNA polymerase can only initiate transcription with the remaining hydroxyl group, thus forcing ARCA incorporation in the forward orientation. As a result, 100% of the transcripts synthesized with ARCA at the 5' end are translatable, leading to a strong stimulatory effect on translation.

Proper capping of in vitro transcribed RNA

Proper capping of RNA promotes correct initiation of protein synthesis, as well as stability and processing of mRNA in vivo. Uncapped RNA is rapidly degraded by cellular RNases after microinjection or transfection into cells. Capped RNA is also typically translated more efficiently in in vitro translation systems, generating RNAs with cap analog incorporated only in the functional orientation. Therefore, substitution of traditional cap analog with ARCA results in the synthesis of capped RNAs that are 100% translatable.

Specifications

• Content And Storage: T7 Enzyme Mix, 10X Reaction Buffer 2XNTP⁄ARCA Solution, GTP Solution, pTRI-Xef, TURBO DNase, Ammonium Acetate Stop Solution, Lithium Chloride Precipitation Solution, E-PAP, 5X E-PAP Buffer, ATP Solution, MnCl₂, and Formaldehyde Load Dye are all stored at –20°C. Nuclease-free Water may be stored at any temperature.
• Final Product Type: Synthetic mRNA, mRNA, ARCA Capped 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: 10 Reactions
• Product Line: Ambion™, mMESSAGE mMACHINE™
• Product Type: T7 In Vitro Transcription kit
• Quantity: 10 Reactions
• Promoter: T7
• Shipping Condition: Dry Ice
• Yield: 20-30 μg
• Starting Material: PCR Products, Linearized DNA plasmid

Safety and Handling

WARNING: Reproductive Harm - www.P65Warnings.ca.gov

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