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Polymerases

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SP6 RNA Polymerase

No Product Name Cat.# Conc Size Price(₩) Add
1 SP6 RNA Polymerase RP003S 20 units/μl 2,000 U ₩ 68,000 추가
2 SP6 RNA Polymerase RP003L 20 units/μl 10,000 U ₩ 272,000 추가
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Description

SP6 RNA Polymerase catalyzes RNA synthesis in the 5´→ 3´ direction. It requires the presence of a DNA template which contains a SP6 phage promoter. SP6 RNA Polymerase can be used for in vitro

translation; RNA probes labeling and prepare mRNA.

 

Characteristics

- Isolated from a recombinant source

- RNA probe preparation for hybridization

- mRNA generation for in vitro translation systems

 

Applications

- Radiolabeled RNA probe preparation

- RNA generation for in vitro translation

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

- Expression control via anti-sense RNA

 

Unit Definition

One unit is defined as the amount of enzyme that will incorporate 1 nmol ATP into acid-insoluble material in a total reaction volume of 50 μl in 1 hour at 40°C in 1X SP6 RNA Polymerase buffer.

 

Storage Conditions

50 mM Tris-HCl, 100 mM NaCl, 20 mM β-ME, 1 mM EDTA, 50% Glycerol, 0.1% Triton® X-100

pH 7.9 @ 25°C, Store at -20°C

 

Quality Control

- Endonuclease -free

- Exonuclease-free

- Non-Specific DNase Activity

- RNA Polymerase Specificity:

- RNase Activity

 

Cautions

- SP6 RNA Polymerase activity depends on dithiothreitol.

- Highly sensitive to salt inhibition. Salt concentration should not exceed 50 mM.

- Higher yields of RNA can be obtained by raising NTP concentrations (up to 4 mM each)

- Reduced enzyme activity over time may be due to the breakdown of dithiothreitol in the reaction

buffer. Add 10 mM fresh dithiothreitol to recover the activity.

 

Standard reaction conditions
- RNA polymerization conditions

10X SP6 RNA Polymerase buffer

μl

SP6 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_RP003_SP6 RNA Polymerase

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