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Image credit: Blair Lyons
Aid mission:
Transcription
Exceptionally high yields of RNA
transcripts in very short time
The Thermo ScientificTM TranscriptAidTM T7 High Yield Transcription
Kit is designed for high yield in vitro transcription from DNA templates
containing a T7 RNA polymerase promoter and is suitable to
synthesize both short (<100 nt) and long transcripts. Depending on
transcript length each reaction yields approximately 200 µg RNA from
1 µg template DNA in 2 hours. For maximum flexibilty, the kit provides
protocols for the generation of unlabeled, labeled and capped RNA.
Besides all reagents for in vitro transcription, loading dye and a RNA
ladder for gel analysis are included with the kit.
Features
• Exceptionally high
yields – up to 200 µg in
2 hours
• Versatile – suitable for
both short and long RNA
transcripts
• Milligram amounts of
RNA in a single, scaledup reaction
Obtained RNA is of high integrity and can be used in numerous
applications including RNA structural and functional studies,
ribozyme biochemistry, RNase protection assays and hybridizationbased blots, anti-sense RNA and RNAi experiments, microarray
analysis, microinjection, in vitro translation, and RNA vaccines related experiments.
• Flexible – generates
unlabeled, labeled or
capped RNA
• High value –
RNA ladder included
 RNA synthesis by in vitro
transcription using linearized
plasmid DNA as a template.
T7
promoter
DNA insert
Linearization
site
L
res inea
tric riza
tio tio
ne nw
nz ith
ym
e
T7
In vitro transcription
with T7 RNA
polymerase
T7
Removal of template
DNA with Dnase I
RNA
transcripts
Product Information
High yields of full length RNA transcripts
Thermo Scientific
Time course of RNA synthesis
for short and long transcripts
M
1
2
3
Vendor 1
4
5
Vendor 2
6
7
8
9
M
250
 RNA transcripts (500 nt, 1000 nt and 6000 nt) were generated using high
yield in vitro transcription reactions containing 1 μg of DNA template. Each
20 μL reaction was incubated for 2 hours at 37°C. The transcription products
were diluted 120-fold and 6 μL was run on a 1% agarose/TAE gel supplemented
with 0.5 μg/mL EtBr.
RNA yield, μg
200
150
M – Thermo ScientificTM RiboRulerTM High Range RNA Ladder, ready-to-use (#SM1823).
1, 4, 7 – 500 nt RNA transcript. 2, 5, 8 – 1000 nt RNA transcript.
3, 6, 9 – 6000 nt RNA transcript.
100
500 nt
1000 nt
6000 nt
50
0
2
High yield of short RNA transcripts
Thermo Scientific
4
Vendor 1
Vendor 2
Time of reaction, h
 RNA transcripts (500 nt, 1000 nt and 6000 nt) were
generated with TranscriptAid T7 High Yield Transcription Kit. 1 μg of DNA template was used in 20 μl
reactions. Yields of RNA were determined at different
time points during the reaction using an Agilent 2100
BioanalyzerTM.
M
3
4
5
6
7
8
9
M
M – Thermo ScientificTM GeneRulerTM Ultra Low Range DNA Ladder, ready-to-use.
1, 4, 7 – 44 nt RNA transcript. 2, 5, 8 – 100 nt RNA transcript. 3, 6, 9 – 200 nt transcript.
200
150
KIT COMPONENTS:
100
TranscriptAid Enzyme Mix
5X TranscriptAid Reaction Buffer
DNase I, RNase-free
ATP, Tris Buffered, 100 mM
CTP, Tris Buffered, 100 mM
GTP, Tris Buffered, 100 mM
UTP, Tris Buffered, 100 mM
44 nt
100 nt
200 nt
50
0
2
6
4
8
Time of reaction, h
 RNA ranscripts (44 nt, 100 nt and 200 nt) were
generated with TranscriptAid T7 High Yield Transcription
Kit. 1 μg of DNA template was used in 20 μL reactions.
RNA yields were determined at different time points
during the reaction using an Agilent 2100 Bioanalyzer.
Control Template (0.5 µg/µL)
3 M Sodium Acetate Solution, pH 5.2
DEPC-treated Water
2X RNA Loading Dye
RiboRuler High Range RNA
Ladder, Ready-to-use
0.5 M EDTA, pH 8.0
Ordering Information
Cat. No
Description
Quantity
K0441
TranscriptAid T7 High Yield Transcription Kit
50 rxns
• For further product information see
thermoscientific.com/onebio
© 2014 Thermo Fisher Scientific Inc. All rights reserved. Bioanalyzer is a registered trademark of Agilent. All other
trademarks are the property of Thermo Fisher Scientific Inc. and its subsidiaries.
MB1418
2
 RNA transcripts (44 nt, 100 nt and 200 nt ) were generated using high yield in vitro
transcription reactions containing 1 µg of DNA template. Each 20 µL reaction was
incubated for 4 hours at 37°C. The transcription products were diluted 120-fold and 10
μL was run on a 5% agarose/TBE gel supplemented with 0.5 μg/mL EtBr.
250
RNA yield, μg
1
Europe
United States
Canada
Customer Service
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Customer Service
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Customer Service
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Technical Support
[email protected]
Technical Support
[email protected]
Technical Support
[email protected]
Tel 00800 222 00 888
Fax 00800 222 00 889
Tel 877 661 8841
Fax 800 292 6088
Tel 800 340 9026
Fax 800 472 8322