RPOLT3-RO
Roche
T3 RNA Polymerase
from Escherichia coli HB101
Synonym(s):
mRNA, polymerase
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About This Item
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biological source
Escherichia coli (HB101)
Quality Level
Assay
100% (SDS-PAGE)
form
solution
specific activity
≥20 U/μL
mol wt
100 kDa (single polypeptide chain)
packaging
pkg of 1,000 U (11031163001)
pkg of 5,000 U (11031171001)
manufacturer/tradename
Roche
concentration
<0.1 % (w/w)
technique(s)
DNA sequencing: suitable
Northern blotting: suitable
Southern blotting: suitable
hybridization: suitable
color
colorless
pH
7.9 (39 °F)
8.0 (68 °F)
solubility
water: miscible
suitability
suitable for molecular biology
NCBI accession no.
UniProt accession no.
application(s)
genomic analysis
life science and biopharma
foreign activity
Nicking activity 150 units, none detected
RNases 150 units
endonucleases 150 units, none detected
storage temp.
−20°C
Gene Information
Escherichia coli ... rpoB(948488)
Related Categories
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.
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.
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
Volume Activity: ≥20 U/μl
Preparation Note
Activator: BSA/spermine
Storage and Stability
Keep container tightly closed in a dry and well-ventilated place.
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
Signal Word
Warning
Hazard Statements
Precautionary 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
Certificates of Analysis (COA)
Search for Certificates of Analysis (COA) by entering the products Lot/Batch Number. Lot and Batch Numbers can be found on a product’s label following the words ‘Lot’ or ‘Batch’.
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Find documentation for the products that you have recently purchased in the Document Library.
Nicole Rosskothen-Kuhl et al.
PloS one, 9(3), e92624-e92624 (2014-03-22)
Brain development and learning is accompanied by morphological and molecular changes in neurons. The growth associated protein 43 (Gap43), indicator of neurite elongation and synapse formation, is highly expressed during early stages of development. Upon maturation of the brain, Gap43
Kathleen Richter et al.
The Journal of biological chemistry, 284(44), 30652-30661 (2009-09-08)
We identified a novel GTPase, SLIP-GC, with expression limited to a few tissues, in particular germinal center B cells. It lacks homology to any known proteins, indicating that it may belong to a novel family of GTPases. SLIP-GC is expressed
Richard Weiszmann et al.
Nature protocols, 4(5), 605-618 (2009-04-11)
We describe a high-throughput protocol for RNA in situ hybridization (ISH) to Drosophila embryos in a 96-well format. cDNA or genomic DNA templates are amplified by PCR and then digoxigenin-labeled ribonucleotides are incorporated into antisense RNA probes by in vitro
Yoshihiro Komatsu et al.
Methods in molecular biology (Clifton, N.J.), 1092, 1-15 (2013-12-10)
In situ hybridization is a powerful method for detecting endogenous mRNA sequences in morphologically preserved samples. We provide in situ hybridization methods, which are specifically optimized for mouse embryonic samples as whole mounts and section tissues. Additionally, β-Galactosidase (β-gal) is
Sandra G Zimmerman et al.
Nature protocols, 8(11), 2158-2179 (2013-10-12)
In situ hybridization (ISH) is a powerful technique for detecting nucleic acids in cells and tissues. Here we describe three ISH procedures that are optimized for Drosophila ovaries: whole-mount, digoxigenin-labeled RNA ISH; RNA fluorescent ISH (FISH); and protein immunofluorescence (IF)-RNA
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