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RT7POL

Sigma-Aldrich

T7 RNA Polymerase

≥50,000 units/mL, aqueous solution

Synonym(s):

bacteriophage T7 RNA polymerase, T7 RNAP, e coli T7 RNA polymerase, in vitro mRNA polymerase, recombinant T7

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About This Item

UNSPSC Code:
12352204

Key Documents

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biological source

bacteriophage T7

Quality Level

200

recombinant

expressed in E. coli

grade

Molecular Biology

Assay

≥95% (SDS-PAGE)

form

aqueous solution (containing 50% glycerol)
liquid

specific activity

≥50000 units/mL

shelf life

2 yr at -20 °C

mol wt

11.4 kDa

storage condition

OK to freeze

concentration

≥50000 units/mL

color

colorless

optimum pH

7.7 (37 °C)

pH

7.9 (25 °C)

suitability

suitable for (In-vitro RNA transcription)

application(s)

research use

foreign activity

DNAse, none detected

RNAse, none detected

Nickase, none detected

shipped in

dry ice

storage temp.

-10 to -25°C

General description

T7 RNA Polymerase is an RNA polymerase from the T7 bacteriophage that catalyzes the formation of RNA from DNA in the 5′ to 3′ direction. The T7 polymerase also requires a double stranded DNA template and Mg2+ ion as cofactor for the synthesis of RNA. It has a very low error rate.

Application

This product is a recombinant protein expressed in E. coli and induced by IPTG. The MW of T7 RNA Polymerase is 98.9 kDa and theoretical pI of ~6.8.

Features and Benefits

T7 polymerase is extremely promoter-specific, transcribes only DNA downstream of a T7 promoter, and can be used for in vitro mRNA synthesis. Quality tests include:
  • Nickase activty: None detected in a 50 µl reaction containing 250 ng of supercoiled pUC19 DNA and at least 30 units of T7 RNA Polymerase incubated for 4 hours at 37°C.
  • DNase Activity: None detected in a 50 µl reaction containing 250 ng linear pUC19 DNA and at least 30 units of T7 RNA Polymerase incubated for 16 hours at 37°C.
  • RNase Activity: None detected in a 50 µl reaction containing 1.25-2.5 ng tRNA and at least 30 units of T7 RNA Polymerase is incubated for 4h at 37°C.

Physical form

The product is supplied as an aqueous solution containing 50mM Tris-HCl, 100mM NaCl, 20mM β-ME, 1mM EDTA, 0.02% (w/v) Tween 20 and 50% glycerol, pH 7.9 at 25 °C.

Other Notes

1 Unit of T7 RNA polymerase is defined as the amount of enzyme that synthesizes 1ug of mRNA per hour in a 30-minute reaction containing 25μg/ml linearized plasmid, 3mM of each NTP and 18mM MgCl2, at 37°C, pH 7.7

Storage Class Code

10 - Combustible liquids

WGK

WGK 2

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Regulatory Information

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Certificates of Analysis (COA)

Lot/Batch Number
0000404475
0000404474
0000404474
0000404475

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Promoter specificity determinants of T7 RNA polymerase.
Rong M, et al.
Proceedings of the National Academy of Sciences of the USA, 95, 515-519 (1998)

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