T7 RNA polymerase is a 99kDa DNA dependent RNA polymerase which initiates RNA synthesis after encountering a T7 phage promoter sequence on the DNA template. This enzyme is essential for in-vitro transcription of a template DNA and hence, is indispensable for the manufacture of mRNA therapeutics and vaccines. We offer high quality T7 RNA polymerase suitable for all research/pre-clinical/clinical applications.

Advantages:

• Manufactured in accordance with GMP rules and regulations
• 100% animal origin free manufacturing process
• Endotoxin and antibiotic free
• Complete QA documentation available

EC Number:

2.7.7.6

Source:

Recombinant E.Coli

Molecular Weight:

34 kD

Cofactor Requirement:

Mg2+ for mRNA synthesis

T7 Promoter Sequence:

Required

Yeast inorganic Pyrophosphatase is expressed with an N-terminal His-Tag in Recombinant E.coli

Appearance:

Clear, colorless solution

Volume Activity:

400KU- 1000 kU/mL

Unit Definition:

One unit is defined as the enzyme activity which incorporates 1 nmol CMP in acid-precipitable RNA within 60 min at +37°C.

Protein (OD280):

0.5-1.2 mg/mL

Purity:

≥ 95 %

Unspecific Nucleases:

Not detectable

Nicking Activity:

Not detectable

RNase Activity:

Not detectable

Exonucleases:

Not detectable

Proteases:

Not detectable

Endotoxins:

NMT 10 EU/mL

Total Bioburden:

NMT 10 CFU/mL

E.coli Host Cell DNA:

NMT 100 pg/mg protein

E.coli Host Cell Protein:

NMT 50 ppm

Heavy Metals:

NMT 10 ppm

Stability:

At -25°C to -15°C within specification range for 18 months from date of manufacture.

Storage:

Enzyme supplied in:

100 mM NaCl, 50 mM Tris-HCl (pH 7.9), 1 mM EDTA, 20 mM 2-mercaptoethanol, 0.1% Triton X-100 and 50% glycerol

Buffer:

10x / 5X buffer will be provided

This reaction mixture works for scale up IVT reactions With 10X buffer (400mM Tris pH 8.0, 100mM DTT, 20mM Spermidine, 0.02% Triton x 100, 165mM Magnesium Acetate) and 5X Reaction buffer ( 400mM HEPES pH 7.5, 200mM DTT, 20mM Spermidine, 120mM MgCl2)

Unit Definition:

One unit is defined as the amount of enzyme required to incorporate 1 nM ATP into an acid-insoluble material in 1 hour at 37°C.

Notes on use:

• For storage of manufactured radio labeled RNA probes, the concentration of the radioactive nucleotide should be limited to 6 μM
• To protect RNA against ribonuclease, an RNase inhibitor should be added to a final concentration of 1 U/μl
• T7 RNA Polymerase is extremely sensitive to salt inhibition. The overall salt concentration should not exceed 50 mM

Method of Analysis will be provided on Bulk Enzyme orders for customer’s in-house quality testing