biotechrabbit | T7 RNA Polymerase, 50 U/µl

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T7 RNA Polymerase, 50 U/µl

Features

High-yield in vitro transcription
Strictly promoter-specific activity
No promoter cross-talk
Free from endonuclease, exonuclease, protease, RNase

Applications

Preparation of homogeneously labeled RNA probes for hybridization (e.g., radioactive, biotin, DIG-11-UTP)
Generation of mRNA for in vitro translation, gene expression or splicing studies
Production of template for cDNA synthesis
In vitro synthesis of capped RNA transcripts
Synthesis of antisense RNA and dsRNA for RNA interference or gene silencing

Purchase T7 RNA Polymerase, 50 U/µl

Availability: In stock Bulk requests please inquire at oem@biotechrabbit.com

Catalog #	Size	Package Content	Price*	Qty
BR1101201	1000 U	20 µl T7 RNA Polymerase 1500 µl 10× T7 RNA Polymerase Reaction Buffer	€60.00
BR1101202	5000 U	100 µl T7 RNA Polymerase 1.5 ml 10× T7 RNA Polymerase Reaction Buffer	€240.00

*Does not include applicable tax, shipping & handling, or other charges.

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biotechrabbit™ T7 RNA Polymerase is an extremely promoter-specific, DNA-dependent polymerase from the T7 bacteriophage. The enzyme requires Mg2+ ions and the presence of the T7 promoter in the template DNA to synthesize RNA in the 5´→ 3´ direction.

The combination of the high-specificity binding, which reduces cross-talk with other promoters, and the rigorous purity testing, which confirms absence of other enzymatic activities, ensures that the T7 RNA Polymerase transcribes large amounts of RNA from sequences downstream of the T7 promoter. By using T7 promoters in opposite orientation, double-stranded RNA can be generated.

The synthesized RNA is suitable for use in a wide range of applications, including hybridization (e.g., blot, in situ, and microarray), RNA protection, capped RNA and antisense RNA.

Component	Composition
T7 RNA Polymerase	T7 RNA Polymerase, 50 U/µl, in storage buffer containing 50% (v/v) glycerol.
T7 RNA Polymerase Reaction Buffer	Optimized 10×T7 RNA Polymerase reaction buffer.

STORAGE

-20°C (until expiry date – see product label)

Unit definition

One unit is the enzyme activity which incorporates 1 nmol CMP in acid-precipitable RNA products within one hour at 37°C.

Quality Control

Exonuclease assay

Linearized lambda/HindII fragments are incubated with T7 RNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Endonuclease assay

lambda DNA is incubated with T7 RNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No degradation of DNA was observed.

Nick Activity

Supercoiled plasmid DNA is incubated with T7 RNA Polymerase in a 50 µl reaction mixture for 4 h at 37°C. No conversion of covalently closed circular DNA to nicked DNA was detected.

RNase Assay

A sample of the enzyme is incubated with a RNA template. RNase activity was not observed after agarose gel.

Prevention of contamination

When assembling the reactions, care should be taken to eliminate the possibility of contamination with undesired RNA, DNA and nucleases.

Use separate clean areas for preparation of samples and reaction mixtures.
Wear fresh gloves. Use sterile tubes and pipette tips with aerosol filters for assay setup.
Use only water and reagents that are free of RNA, DNA and nucleases.
With every assay setup, perform a contamination control reaction that does not include a template.

Basic Protocol

Component	Volume/concentration
10× T7 RNA Polymerase Buffer	10 µl
Nucleotides (ATP, GTP, CTP, UTP)	3.75 mM each
RNAse Inhibitor (optional)	100 U
Template DNA	4 μg
T7 RNA Polymerase (50 U/µl)	2–4 µl
Nuclease free water	add up to 100 µl
Total volume	100 µl