All products have special prices for bulk purchase, please contact for more details if required.

Cat. No.: SP6-1k (for 1KU)

Cat. No.: SP6-5k (for 5KU)

Cat. No.: SP6-25k (for 25KU)

Cat. No.: SP6-100k (for 100KU)

Description

SP6 RNA Polymerase is a DNA-dependent 5'→3' RNA polymerase that exhibits high specificity for the SP6 promoter sequence. It catalyzes the incorporation of NTPs downstream of the SP6 promoter, whether in single-stranded or double-stranded DNA, to synthesize RNA complementary to the template DNA downstream of the SP6 promoter.

Features

SP6 RNA Polymerase is not only capable of regular RNA synthesis but also recognizes modified NTPs, such as biotin-labeled, digoxigenin-labeled, and luciferin-labeled NTPs, enabling the synthesis of labeled RNA for various applications. It exhibits a high degree of specificity for the SP6 promoter.

Applications

Used for RNA synthesis. The synthesized RNA can be used for or as: hybridization probes, genomic DNA sequence analysis, RNase protection assay, antisense RNA synthesis, RNA templates for in vitro translation, substrates for RNA splicing studies, RNA secondary structure and RNA-protein interactions, nucleic acid amplification analysis, siRNA, miRNA, and other small RNAs.

Source

Expressed in Escherichia coli, with the expression gene derived from bacteriophage SP6 RNA Polymerase.

Activity Definition

The amount of enzyme required to catalyze the incorporation of 1 nmol of AMP into a polymeric nucleotide in 60 minutes at 37°C is defined as 1 unit of activity.

Activity Assay Conditions

40 mM Tris-HCl (pH 8.0), 6 mM MgCl2, 10 mM DTT, 2 mM spermidine, 0.5 mM NTP, 0.6 MBq/ml [3H]-ATP, 20 µg/ml plasmid DNA containing the specific SP6 RNA Polymerase promoter sequence.

Purity

Free of DNA endonucleases, exonucleases, and RNases.

Enzyme Storage Solution

50 mM Tris-HCl (pH 7.9, 25°C), 100 mM NaCl, 5 mM DTT, 1 mM EDTA, 50% (v/v) Glycerol, 0.1% (w/v) Triton X-100.

Transcription Buffer (10X)

400 mM Tris-HCl (pH 7.9, 25°C), 20 mM spermidine, 60 mM MgCl2, 10 mM DTT.

Inactivation or Inhibition

SP6 RNA Polymerase can be inactivated by heating at 70°C for 10 minutes. The addition of an appropriate amount of EDTA can also deactivate SP6 RNA Polymerase. Chelating agents and sodium, potassium, or ammonium salts with concentrations greater than 150 mM can significantly inhibit the activity of SP6 RNA Polymerase.

The SP6 consensus promoter sequence is as follows, with the G indicating the transcription start site:

ATTTAGGTGACACTATAGAAGNG

Precautions

• The enzyme should be stored in a cooler or ice bath during use and immediately placed at -20°C after use.
• This product is intended for scientific research use by professionals only. It is not intended for clinical diagnosis or treatment, and should not be used for food or drugs. It should not be stored in a regular household setting.
• For your safety and health, please wear appropriate laboratory attire and disposable gloves when handling.

Storage

Store at -20°C.