RPOLT3-RO

T3 RNA Polymerase

from Escherichia coli HB101

• Synonyme(s) : mRNA, polymerase
• Numéro de classification (Commission des enzymes): 2.7.7.6 (BRENDA, IUBMB)
• Code UNSPSC : 12352204

Propriétés

• Source biologique: Escherichia coli (HB101)
• Niveau de qualité: 100
• Pureté: 100% (SDS-PAGE)
• Forme: solution
• Activité spécifique: ≥20 U/μL
• Poids mol.: 100 kDa (single polypeptide chain)
• Conditionnement: pkg of 1,000 U (11031163001); pkg of 5,000 U (11031171001)
• Fabricant/nom de marque: Roche
• Concentration: <0.1 % (w/w)
• Technique(s): DNA sequencing: suitable; Northern blotting: suitable; Southern blotting: suitable; hybridization: suitable
• Couleur: colorless
• pH: 7.9 (39 °F); 8.0 (68 °F)
• Solubilité: water: miscible
• Adéquation: suitable for molecular biology
• Numéro d'accès NCBI: NP_418414
• Numéro d'accès UniProt: P0A8V2
• Application(s): genomic analysis; life science and biopharma
• Activité étrangère: Nicking activity 150 units, none detected; RNases 150 units; endonucleases 150 units, none detected
• Température de stockage: −20°C
• Informations sur le gène: Escherichia coli ... rpoB(948488)

Description

Description générale

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.

Spécificité

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.

Conditionnement

1 kit containing 2 components

Qualité

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

Caractéristiques

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.

Définition de l'unité

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

Notes préparatoires

Activator: BSA/spermine

Stockage et stabilité

Keep container tightly closed in a dry and well-ventilated place.

Autres remarques

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

Composants de kit seuls

• T3 RNA Polymerase, in buffer, pH 7.9 ≥20 U/μl
• Transcription Buffer 10x concentrated

Informations sur la sécurité

• Pictogrammes: GHS07
• Mention d'avertissement: Warning
• Mentions de danger: H319
• Conseils de prudence: P264 - P280 - P305 + P351 + P338 - P337 + P313
• Classification des risques: Eye Irrit. 2
• Code de la classe de stockage: 12 - Non Combustible Liquids
• Classe de danger pour l'eau (WGK): WGK 2
• Point d'éclair (°F): does not flash
• Point d'éclair (°C): does not flash