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:3/29/2017)... , March 29, 2017 /PRNewswire/ -  GeneNews Limited ... BreastSentry™ , a new risk stratification test for breast ... lab, Innovative Diagnostics Laboratory ("IDL"). BreastSentry incorporates a blood-based biomarker ... and lifetime risk for developing breast cancer.   ... BreastSentry measures the ...
(Date:3/29/2017)... , March 29, 2017 The Global ... 2022 report is a specialized and comprehensive study on the existing ... North America , Europe and ... Middle East and Africa . ... Browse 172 Tables ...
(Date:3/28/2017)... ... March 28, 2017 , ... NetDimensions announced today ... and enhance training plan management for consistent implementation of standards and regulatory requirements ... SHL Group to help improve and streamline their training and employee development programs, ...
(Date:3/28/2017)... ... March 28, 2017 , ... Franz Inc ., the leading ... , has been named a ‘Champion’ by Bloor Research in its recent Graph ... in its class, and, thanks to Gruff, it was rated as the easiest product ...
Breaking Biology Technology:
(Date:3/9/2017)... Australia , March 9, 2017 /PRNewswire/ ... at the prestigious World Lung Imaging Workshop at the ... Fouras , was invited to deliver the latest data ... This globally recognised event brings together leaders at the ... latest developments in lung imaging. "The ...
(Date:3/7/2017)... BRIGHTON, England , March 7, 2017 Brandwatch ... been chosen by The Prince,s Trust to uncover insights ... insights across The Trust. The UK,s leading youth charity ... track social campaign results and get a better understanding of the ... ...
(Date:3/6/2017)... , March 6, 2017 Mintigo ... technology, today announced Predictive Sales Coach TM , ... actionable sales intelligence into Salesforce. This unique AI ... their sales organizations with deep knowledge of their ... for intelligent engagement. Predictive Sales Coach extends Mintigo,s ...
Breaking Biology News(10 mins):