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capped RNA
dsRNA transcripts
uncapped RNA
amplified RNA
short RNA transcripts
milligram quantities of transcript
MEGAscript: Consistent, High Yields of RNA Transcripts
Using the MEGAscript
Kit, you can produce over 120 g of RNA from 1 g of DNA template.
This is 10-50 times the amount of RNA produced by conventional transcription
(Figure 2). Reactions are versatile and can be manipulated to include
specialized reagents such as modified nucleotides or additional polymerase.
In addition, Ambion has optimized the transcription of short (20-500 nt)
DNA templates with the MEGAshortscript
Kit. This patented technology was designed to maximize the yield of
short RNA transcripts from plasmid, oligonucleotides, and PCR product
templates.
Figure 1. Electropherogram of RNA Transcripts of Different Sizes. In vitro transcription was performed using 1, 4, or 9 kb templates with Ambion's MEGAscript Kit according to protocol. 20 l reactions were diluted to 100 l and 1 l was assessed on the Agilent 2100 bioanalyzer. The single primary peak is indicative of product quality while the area under the peak is related to yield.
Figure 2. MEGAscript vs. Conventional In Vitro Transcription Reactions. 20 l transcription reactions containing 1 g template, T7 MEGAscript components and [32P]UTP, were incubated at 37C. 1 l aliquots were removed at 1 hr intervals and acid precipitable cpm determined by scintillation counting.
mMESSAGE mMACHINE: Translatable Capped RNA
For high yields of capped RNA, MEGAscript technology
is incorporated into the mMESSAGE
mMACHINE Kit. This kit contains an optimized ratio of cap analog
to GTP, for the best trade off between yield and proportion of capped
transcripts (80% capped). Furthermore, you can obtain 100% translatable
capped RNA with the mMESSAGE
mMACHINE T7 ULTRA Kit. This kit uses a unique cap analog that only
incorporates in the functional, translatable orientation, resulting in
the synthesis of capped RNAs that are 100% translatable.
MEGAscript RNAi: dsRNA Synthesis for
RNAi
In one complete kit, you
can generate ready-to-use double-stranded RNA (dsRNA) for RNAi
experiments in non-mammalian systems. Unlike competing
products, the MEGAscript
RNAi Kit provides reagents to eliminate DNA template,
single stranded RNA, and free nucleotides. The MEGAscript RNAi
Kit allows researchers to synthesize 50 to 80 g or more of
dsRNA, up to 1.2 kb in length, in a single transcription
reaction.
MessageAmp II: RNA Amplification
Amplify RNA with MEGAscript technology using the
MessageAmp
II aRNA Amplification Kit. The MessageAmp II Kit can amplify as little
as 100 ng of input RNA in a single round for GeneChip analysis. A rationally
engineered M-MLV RT, ArrayScript,
is provided in the kit to maximize yields of full-length cDNA. cRNA synthesis
is powered by MEGAscript technology. These features result in high labeling
efficiency, yielding consistently high percentages of Present Calls, and
low 3'/5' ratios in GeneChip analysis. This procedure is compatible with
biotin, cyanine dye (Perkin Elmer), and CyDye (Amersham Biosciences)
labeling either during or after aRNA amplification. For additional information,
see RNA Amplification
Just Got Better.
Bulk Transcription-- Milligram Quantities
of RNA
If you have a project
or application that requires large amounts of RNA transcripts,
Ambion can meet your needs with its bulk and custom products.
Our MEGAscript technology has been optimized to produce
milligram quantities of RNA. Contact custom@ambion.com for bulk
orders or inquiries.
Ordering Information
Cat#
Product Name
Size
1330
MEGAscript SP6 Kit
40 rxns
1333
MEGAscript T7 Kit
25 rxns
1334
MEGAscript T7 Kit
40 rxns
1338
MEGAscript T3 Kit
40 rxns
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
1354
MEGAshortscript T7 Kit
25 rxns
1626
MEGAscript RNAi Kit
20 rxns
1751
MessageAmp II aRNA
Amplification Kit
20 rxns
B1334-5
5X MEGAscript T7 Kit
200 rxns
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