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Rapidly Synthesize Large Amounts of Capped RNA

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 (

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