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mMESSAGE mMACHINE® T7 ULTRA Transcription Kit

日期: 2015-06-01
浏览次数: 351

货号: AM1345

 

规格

 

Starting Material:Linearized DNA, PCR Products
Promoter:T7
RNA Yield:20-30 µg
Final Product:In Vitro Transcribed RNA, ARCA Capped RNA
Product Size:10 reactions
Shipping Condition:Dry Ice

 

内容及储存

 

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.

 

描述

 

The mMESSAGE mMACHINE® T7 Ultra Kit combines a new cap analog, 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. Advantages of the mMESSAGE mMACHINE® T7 Ultra Kit:

• 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.

 

 

 

 

 

 

 

 

 

 

 

 

 

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