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RPOLT3-RO

Roche

T3 RNA Polymerase

from Escherichia coli HB101

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Synonym(s):
mRNA, polymerase
Enzyme Commission number:

Quality Level

form

solution

packaging

pkg of 1,000 U (11031163001)
pkg of 5,000 U (11031171001)

manufacturer/tradename

Roche

storage temp.

−20°C

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This Item
RPOLT7-RO1148002200105-952-I
T3 RNA Polymerase from Escherichia coli HB101

Roche

RPOLT3-RO

T3 RNA Polymerase

T7 RNA Polymerase from Escherichia coli BL 21/pAR 1219

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Tth DNA Polymerase pkg of 500 U (2 x 250_U), sufficient for ≤200 reactions

Roche

11480022001

Tth DNA Polymerase

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

Roche

manufacturer/tradename

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

-

Quality Level

100

Quality Level

100

Quality Level

100

Quality Level

-

packaging

pkg of 1,000 U (11031163001)

packaging

pkg of 1,000 U (10881767001), pkg of 5,000 U (10881775001)

packaging

pkg of 500 U (2 x 250_U)

packaging

antibody small pack of 25 μg

General description

With this enzyme, homogeneously labeled single-stranded RNA can be generated. Transcripts can be nonradioactively labeled with biotin or DIG-11-UTP or radioactively labeled to high specific activity with [α-32P]- or [α-35S]-labeled nucleotides. T3 RNA polymerase is a DNA-dependent RNA polymerase that is highly specific for its promoter. Only DNA downstream from the T3 promoter will be transcribed.

Specificity

Promotor specifity
T3 RNA polymerase is extremely promoter-specific and only transcribes bacteriophage T3 DNA or DNA cloned downstream of a T3 promoter.
Heat inactivation: Stop the reaction by adding 2 μl 0.2 M EDTA (pH 8.0) and/or heat to 65 °C.

Application

T3 RNA Polymerase can transcribe RNA from cloned DNA templates that are downstream from a T3 promoter. The synthesis can be performed with labeled NTPs to generate highly labeled RNA. Synthesized RNA can be used in many applications, including:
  • RNA or DNA blotting techniques
  • In situ hybridization
  • RNase protection studies: Transcripts synthesized by the enzyme are used as precursor RNA for studies on RNA splicing and processing.
  • Synthesis of capped RNA in vitro with addition of m7GpppG or m7GpppA in excess over GTP or ATP during the transcription reaction. The generated antisense RNA can be introduced into cells to suppress the expression of the corresponding genes.

Packaging

1 kit containing 2 components

Quality

Absence of endonucleases, nicking activity, and RNases tested according to the current Quality Control procedures. Function tested with the SP6/T7 Transcription Kit.

Specifications

Volume Activity: =20U/μl
Synthesis of hybridization probes: T3 RNA polymerase allows highly efficient production of homogeneously labeled RNA. This labeled RNA may be used as hybridization probes in Southern, northern, and dot blots, as well as in situ hybridizations.
Suitable labels: Transcripts can be nonradioactively labeled with biotin-16-UTP, DIG-11-UTP, or fluorescein-12-UTP. They may also be radioactively labeled to high specific activity with [a-32P]- or [a-35S]-labeled nucleotides.

Note: Roche has 10x concentrated RNA Labeling Mixes that are specially designed for DIG-, biotin-, and fluorescein-labeling. These mixes work well with T3 RNA Polymerase.

Unit Definition

One unit is the enzyme activity which incorporates 1 nMol CMP in acid-precipitable RNA products within 60 minutes at +37 °C.

Volume Activity: ≥20 U/μl

Preparation Note

Activator: BSA/spermine

Other Notes

For life science research only. Not for use in diagnostic procedures.

Kit Components Only

Product No.
Description

  • T3 RNA Polymerase, in buffer, pH 7.9 ≥20 U/μl

  • Transcription Buffer 10x concentrated

Pictograms

Exclamation mark

Signal Word

Warning

Hazard Statements

Hazard Classifications

Eye Irrit. 2

Storage Class Code

12 - Non Combustible Liquids

WGK

WGK 2

Flash Point(F)

does not flash

Flash Point(C)

does not flash


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