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T7 RNA Polymerase

No Product Name Cat.# Conc Size Price(₩) Add
1 T7 RNA Polymerase RP001S 50 units/μl 5,000 units ₩ 80,000 추가
2 T7 RNA Polymerase RP001M 50 units/μl 10,000 units ₩ 144,000 추가
3 T7 RNA Polymerase RP001L 50 units/μl 25,000 units ₩ 320,000 추가
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Product description

T7 RNA Polymerase is produced by expressing the RNA polymerase gene of bacteriophage T7 in   E. coli, and purified to homogeneity. T7 RNA Polymerase binds specially to the T7 promoter and synthesizes RNA transcripts very efficiently.

 

Characteristics

- Molecular weight: 98 kDa

- Reaction temperature: 37

- Thermal stability: Half life of 2 min at 50

- Heat inactivation: 70, 10 min

- Active only with the double-stranded T7 promoter sequence.

 

Applications

- Preparation of radioisotope-labeled RNA probes

- RNA synthesis for in vitro translation

- RNA synthesis for research on RNA structure, processing, and catalysts

- Preparation of anti-sense RNA for studies of regulation of gene expression

 

Supplied with

- 10X T7 RNA Polymerase buffer

- 10X DTT

- Sterile water (RNase free)

 

Quality control

- Purity: >99% on SDS-PAGE

- Endonuclease-free

- Exonuclease-free

- RNase-free

 

Unit definition

One unit is defined as the amount of T7 RNA Polymerase required to incorporate 1 nmol of ATP into acid-insoluble materials in 1X T7 RNA Polymerase buffer in 1 hr at 37 with DNA contained double-stranded T7 promoter sequence (1 μg) as template.

 

Storage buffer

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

 

10 X T7 RNA Polymerase buffer

400 mM Tris-HCl (pH 7.9), 250 mM MgCl2, 20 mM Spermidine

 

Cautions

- T7 RNA Polymerase requires dithiothreiotol (DTT) for activity. (Long-term storage may cause oxidation of DTT, 

  and addition of freshly-prepared DTT can reactivate the enzymatic activity)

- Total concentration of salt (such as NaCl or KCl) should not exceed 50 mM for the optimal result.

 

Standard reaction conditions
- RNA polymerization conditions

10X T7 RNA Polymerase buffer

μl

T7 RNA polymerase (50 unit/μl)

μl

rNTP mixture (5 mM each)

μl

10X DTT

5 μl

Double-stranded DNA template (1 μg/μl)

1 μl

RNase Inhibitor (40 unit/μl)

1 μl

Sterile water (RNase free)

up to 50 μl

→Incubate at 37 for 60 to 120 min

→Terminate reaction by incubating at 75 for 20 min, or adding 2 μl of 0.5M EDTA

Reagents and materials not provided: rNTP, RNase Inhibitor 

● Material Safety Data Sheet

- Enzynomics_MSDS_RP001_T7 RNA Polymerase

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