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T7
RNA Polymerase (Lyophilized)

T7 RNA Polymerase is a highly processive enzyme that shows strict specificity for its double stranded promoter sequence (5´ TAATACGACTCACTATAG 3´) and catalyzes the 5’ - 3’ synthesis of RNA.

The T7 RNAP has been obtained from E. coli BL21 cells with a cloned gene encoding the enzyme and purified using our ArtPure™ proprietary purification process, yielding a >99% pure, animal origin-free product that is free of contaminants such as proteinases, RNases and DNases and has no added enzyme inhibitors.

Our T7 RNA Polymerase has been lyophilized in a stable form, ensuring integrity during transport while significantly reducing shipping complexity and costs. For eco-friendly, reliable, cost-effective, and convenient handling, our lyophilized T7 RNA Polymerase is the superior choice.

IVT PROTOCOL

Suggested IVT protocol for optimized ArtPure™ T7 activity

ENZYME QUALITY

ArtPure™ T7 is free of contaminants ensuring high yield of pure RNA

PRODUCT INFORMATION

Optimal storage conditions ensure prolonged enzyme function

ORDER

Get in touch to get more information and a quote

Suggested IVT Protocol

Assemble the following reaction at room temperature.

The volumes can be scaled according to the application.

Component
Amount
Linearized DNA template
500 ng
Transcription buffer (10X)
1 µl
NTP
1 mM each
RNase inhibitor
1 U/µl
T7 RNA Polymerase
0.2 µl
Nuclease-free water
to 10 µl

Incubate the reaction 1 hour at 37°C.

To remove the DNA template, add 1 U of DNase I and incubate at 37°C for 15 minutes.

Inactivate the DNase I by adding 5 mM of EDTA, incubate at 70°C for 10 minutes.

Enzyme Quality

Purity

T7_lyo_purity.png

RNases

T7_lyo_RNases.png

Activity

T7_lyo_activity.png

Proteinases

T7_lyo_proteases.png

DNases

Single-stranded exonucleases

Double-stranded endonucleases

Double-stranded exonucleases

    0h        4h       16h       0h       4h       16h         0h        4h       16h    

T7_lyo_nucleases.png

Definitions

Activity (U/µl): One enzyme unit incorporates 1 nmol of soluble RNA nucleotides into insoluble polymeric RNA per hour at 37°C.

Purity: Evaluated by SDS-PAGE, where the integrated peak area of the RNA Polymerase is compared to other protein peaks in the same lane.

Transcriptional activity: Quantification of total RNA products after in vitro transcription and RNA purification.

 

Proteinases: Assessed using Pierce™ Fluorescent Protease Assay Kit (ThermoFisher Scientific) following the manufacturer’s guidelines.

RNases: Assessed using the RNAse Alert kit (Integrated DNA Technologies), following the manufacturer’s guidelines. 

Single-stranded exonucleases: 2.5 µl of the T7 RNAP are incubated with 0.5 µg of ssDNA substrate (calf-thymus DNA) for 0 hours, 4 hours at 37°C and 16 hours at room temperature.

Double-stranded endonucleases: 2.5 µl of the T7 RNAP are incubated with 0.5 µg of supercoiled plasmid DNA for 0 hours, 4 hours at 37°C and 16 hours at room temperature.  

Double-stranded exonucleases: 2.5 µl of the T7 RNAP are incubated with 0.5 µg of linearized double-stranded DNA substrate (DNA ladder) for 0 hours, 4 hours at 37°C and 16 hours at room temperature.  

Certificates of Analysis

Product Information

Enzyme reconstitution: The enzyme should be reconstituted with the provided buffer upon receipt. To resuspend the lyophilized T7 RNA Polymerase, add 100 µl of the supplied storage buffer to the bottom of the tube. Avoid touching the pellet with the pipette tip. Incubate 5 minutes on ice, or until the buffer has been completely absorbed. Mix by carefully pipetting several times. Do not vortex. Note that modifying the resuspension volume might affect RNA yields.

Storage upon receipt: Our T7 RNA Polymerase is stable at -20°C for up to 12 months in the glycerol-based supplied buffer. For extended storage, we recommend aliquoting in smaller volumes, freezing at -80°C and avoiding repeated freeze-thaw cycles.

Storage buffer: 50 mM Tris-HCl, 100 mM NaCl, 20 mM DTT, 1 mM EDTA, 50% glycerol, 0.1% Triton X-100, pH 7.9. This buffer is shipped at ambient temperature. We recommend storing it at -20°C upon receipt.

Transcription buffer (10X): 400 mM Tris-HCl, 60 mM MgCl2, 100 mM DTT, 100 mM NaCl, 20 mM spermidine, pH 7.9.

Order

Product
SKU
Pack size
Price
T7 RNA Polymerase (Lyophilized)
Z0721
10000 U
T7 RNA Polymerase (Lyophilized)
Z0723
30000 U
T7 RNA Polymerase (Lyophilized)
Z072X
Custom
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