Navigation Links
Rapidly Synthesize Large Amounts of Capped RNA


Convenient format - premixed cap analog and nucleotides provided

Optimized for highest yield of capped RNA

80% capping efficiency

Complete with SUPERaseIn RNase inhibitor, DNase I, and positive control

Ideal for microinjection, in vitro or in vivo translation


The mMESSAGE mMACHINE High Yield Capped RNA Transcription Kits use Ambion's patented high yield transcription technology. Each reaction synthesizes 10-50 times the yield of capped RNA obtained with conventional in vitro transcription reactions.

The reaction is simple. DNA template is mixed with the reaction components and incubated at 37C for up to 2 hours. Figure 1 shows a time course experiment that plots yield of a 3.7 kb capped transcript over time. With this template, the mMESSAGE mMACHINE reaction is complete in only 30 minutes. In addition to synthesizing high yields of capped RNA transcripts (Figure 1), mMESSAGE mMACHINE reactions also produce full-length transcripts. Figure 2 shows an Agilent bioanalyzer 2100 electropherogram of the 3.7 kb transcript. The presence of a single, strong peak indicates the predominance of full length transcript. This is also evident in Figure 3, in which mMESSAGE mMACHINE reactions producing 1.7 kb, 3.7 kb and 9 kb transcripts were analyzed on a conventional denaturing agarose gel.

Figure 1. mMESSAGE mMACHINE Reaction Time Course. A 3.7 kb template was transcribed in 3 replicate mMESSAGE mMACHINE reactions as per protocol. After the indicated incubation times, each reaction was DNase treated as per protocol and a preliminary quantitatation was performed with RiboGreen (Molecular Probes). Each reaction was then purified using the MEGAclear Kit (Ambion) and quantitated by UV spectrophotometry.

Figure 2. Electropherogram of a mMESSAGE mMACHINE Reaction. A 3.7 kb template was used in a 20 l mMESSAGE mMACHINE reaction as per protocol and incubated for 30 min followed by analysis using the Agilent 2100 bioanalyzer.

Figure 3. Gel Analysis of mMESSAGE mMACHINE Reactions. Templates generating transcripts of sizes 1.7 kb, 3.7 kb and 9 kb were used in a mMESSAGE mMACHINE reaction as per protocol. One g of each reaction was denatured using NorthernMax-Gly Sample Loading Dye and analyzed on a 1% agarose gel with EtBr.


Cap analog is premixed with GTP at a ratio optimized for the best compromise between yield and proportion of capped transcripts (80%). The cap analog:GTP ratio can be adjusted with the GTP Solution provided in the kit to increase yields of long transcripts (>4 kb); in fact, RNA transcripts as long as 23 kb have been synthesized with mMESSAGE mMACHINE. The kit includes SUPERase In RNase Inhibitor . SUPERase In inhibits RNases A, B, C, 1 and T1 and offers the greatest available protection against RNases (see data). DNase I and a positive control DNA template are also included. The RNA synthesized from a mMESSAGE mMACHINE reaction is ideal for in vitro or in vivo translation studies.

Ambion also offers the mMESSAGE mMACHINE T7 Ultra Kit for the generation of RNA transcripts capped with Anti Reverse Cap Analog (ARCA). A base modification in this cap analog generates RNAs with ARCA incorporated only in the functional, translatable orientation, resulting in higher protein yields. The mMESSAGE mMACHINE T7 Ultra Kit includes reagents for both transcription and the addition of a poly(A) tail to capped transcripts.


back to top


Ordering Information
Cat# Product Name Size 1340 mMESSAGE mMACHINE SP6 Kit 25 rxns 1344 mMESSAGE mMACHINE T7 Kit 25 rxns 1345 mMESSAGE mMACHINE T7 ULTRA Kit 10 rxns 1348 mMESSAGE mMACHINE T3 Kit 25 rxns
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. Fluorescence-Based Single-Tube Assays to Rapidly Detect Human Gene Mutations
2. HSVision Molecular Beacon Detection Module Rapidly Detects Herpes Simplex Virus DNA
3. Rapidly Prepare Probes and Markers for Small RNA Analysis
4. Synthesize High Yields of Biotinylated aRNA
5. High-Efficiency Site-Directed Mutagenesis of Large Plasmid Vectors
6. PCR-Grade Nucleotides Larger Pack Sizes Now Available
7. Efficient Cloning and Electro-transformation of Large Eukaryotic DNA Fragments
8. Sub-Micron Gold Particles Are Superior to Larger Particles for Efficient Biolistic Transformation of Organelles and Some Cells
9. Constructing Directional cDNA Libraries from Limited Amounts of RNA
10. Detecting Attomole Amounts of Small RNA
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/23/2016)... , June 23, 2016 A person commits ... the crime scene to track the criminal down. ... U.S. Food and Drug Administration (FDA) uses DNA evidence to ... Sound far-fetched? It,s not. The FDA has ... to support investigations of foodborne illnesses. Put as simply as ...
(Date:6/23/2016)... ... June 23, 2016 , ... Charm Sciences, ... microbial test has received AOAC Research Institute approval 061601. , “This is another ... year,” stated Bob Salter, Vice President of Regulatory and Industrial Affairs. “The Peel ...
(Date:6/23/2016)... , June, 23, 2016  The Biodesign Challenge ... envision new ways to harness living systems and biotechnology, ... Art (MoMA) in New York City ... 130 participating students, showcased projects at MoMA,s Celeste Bartos ... Paola Antonelli , MoMA,s senior curator of architecture and ...
(Date:6/23/2016)... ... June 23, 2016 , ... Supplyframe, the Industry ... Supplyframe Design Lab . Located in Pasadena, Calif., the Design Lab’s mission ... hardware projects are designed, built and brought to market. , The Design Lab ...
Breaking Biology Technology:
(Date:4/28/2016)... , April 28, 2016 First quarter ... (139.9), up 966% compared with the first quarter of 2015 ... totaled SEK 589.1 M (loss: 18.8) and the operating margin was ... (loss: 0.32) Cash flow from operations was SEK 249.9 ... 2016 revenue guidance is unchanged, SEK 7,000-8,500 M. The ...
(Date:4/19/2016)... , UAE, April 20, 2016 ... implemented as a compact web-based "all-in-one" system solution for ... biometric fingerprint reader or the door interface with integration ... modern access control systems. The minimal dimensions of the ... readers into the building installations offer considerable freedom of ...
(Date:4/14/2016)... 14, 2016 BioCatch ™, ... today announced the appointment of Eyal Goldwerger ... Goldwerger,s leadership appointment comes at a time ... the deployment of its platform at several of the ... which discerns unique cognitive and physiological factors, is a ...
Breaking Biology News(10 mins):