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TOP-T7 RNA Polymerase

No Product Name Cat.# Conc Size Price(₩) Add
1 TOP-T7 RNA Polymerase RP002S 50 units/μl 5,000 units ₩ 100,000 추가
2 TOP-T7 RNA Polymerase RP002M 50 units/μl 10,000 units ₩ 180,000 추가
3 TOP-T7 RNA Polymerase RP002L 50 units/μl 25,000 units ₩ 400,000 추가
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Product description

TOP-T7 RNA Polymerase is expressed and purified from E. coli to near homogeneity. This product has increased thermostability. It can utilize the T7 promoter in double stranded DNA to transcribe a gene located downstream of the promoter.

 

Characteristics

- Molecular weight: 98 kDa

- Reaction temperature: 45

- Thermal stability: Half life of 84.5 min at 50

- High specificity to T7 promoter sequence in double-stranded DNA

 

Applications

- Preparation of radioisotope-labeled RNA probe

- RNA synthesis for in vitro translation

- RNA synthesis for studies of RNA structure, RNA processing, and RNA catalysis

- Preparation of anti-sense RNA for gene expression studies

 

Supplied with

- 10X TOP-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 TOP-T7 RNA Polymerase required to incorporate 1 nmol of ATP into acid-insoluble materials in 1X TOP-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-HC (pH 7.9), 100 mM NaCl, 20 mM β-mercaptoethanol, 1 mM EDTA, 0.1% Triton X-100, 50% glycerol

 

10X TOP-T7 RNA Polymerase buffer

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

 

Cautions

- DTT is essential for TOP-T7 RNA Polymerase activity. (Long-term storage may cause oxidation of

DTT, resulting in loss of activity. In this case, addition of freshly prepared DTT can restore TOP-T7

RNA Polymerase activity).

- The total concentration of salt should not exceed 50 mM.

 

Standard reaction conditions
- RNA polymerization conditions

10X TOP-T7 RNA Polymerase buffer

μl

TOP-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_RP002_TOP-T7 RNA Polymerase

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