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High Yield Transcription for Every Application
From the Recognized Leader in High Yield
Transcription
The MEGAscript® Kit
is the most frequently cited large-scale transcription kit,
and with good reason. It enables the consistent transcription
of high yields of full-length RNA (Figure 1). Ambion offers
a complete line of products specifically optimized to generate
the following products:
•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 µg of RNA from 1 µg 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.
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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 µl reactions were diluted
to 100 µl and 1 µl 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.
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Figure 2. MEGAscript® vs.
Conventional In Vitro Transcription Reactions. 20 µl
transcription reactions containing 1 µg template,
T7 MEGAscript components and [32P]UTP, were
incubated at 37°C. 1 µl aliquots were removed
at 1 hr intervals and acid precipitable cpm determined
by scintillation counting.
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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 µg 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 Cy™Dye (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.
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