产品名称
RNA聚合酶 来源于肠沙门氏菌 肠道血清型，SP6-感染, buffered aqueous solution
grade
Molecular Biology
form
buffered aqueous solution
concentration
10000-50000 units/mL
UniProt accession no.
foreign activity
DNase, RNase, protease, none detected
nonspecific endonuclease, absence of activity (after 20 hour incubation)
storage temp.
−20°C
Gene Information
bacteriophage SP6 ... SP6p08(1481778)
General description
Sp6 Polymerase is a DNA-dependent RNA polymerase with high specific activity for the bacteriophage Sp6 promoter. The enzyme is efficient in transcribing large quantities of RNA from the DNA sequence (polylinker) downstream from the promoter. Simultaneous transcription by Sp6 and Sp7 polymerases may occur without interfering with each other.
Other Notes
One unit will catalyze the incorporation of 1 nmol of rNTP into acid-precipitable material in 60 min at 37 °C.
Protease is supplied in a solution of 100 mM NaCl, 50 mM Tris-HCl (pH 7.9), 0.1 mM EDTA, 0.1% Triton X-100, 1 mM DTT, and 50% (v/v) glycerol.
Sp6 polymerase exhibits 30% more activity at 40°C than at 37°C.
Analysis Note
Activity assay: 40 mM Tris-HCl, pH 7.9, 6 mM MgCl2, 4 mM spermidine, 10 mM DTT, 0.5 μM each rNTP + 10 μCi α-32P-UTP, 3-10 units of enzyme, and 1 μg of a 350 bp template are incubated for 10 min at 37 °C in a total volume of 100 μl. Typical results are ≥50% incorporation of labeled nucleotide into ≥90% full-length transcript.
Application
Suitable for:
- Synthesis of labeled and unlabeled RNA transcripts
- RNase protection assays
- Generating RNA transcripts for in vitro translation
- Use in in vitro transcription of RNA transcripts, antisense RNA (from reversed cloned DNA insert), and biologically active mRNA
signalword
Warning
hcodes
Hazard Classifications
Aquatic Chronic 3 - Eye Irrit. 2
存储类别
10 - Combustible liquids
wgk
WGK 2
flash_point_f
Not applicable
flash_point_c
Not applicable
法规信息
新产品
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Sambrook, J., et al.
Molecular Cloning: A Laboratory Manual, 18-18 (1989)
M R Green et al.
Cell, 32(3), 681-694 (1983-03-01)
To study the mechanisms of RNA splicing we have synthesized beta-globin mRNA precursors by in vitro transcription using a plasmid in which a human beta-globin gene is fused to an efficient bacteriophage promoter. The structural requirements for accurate splicing of
E T Schenborn et al.
Nucleic acids research, 13(17), 6223-6236 (1985-09-11)
The in vitro synthesis of extraneous RNA sequences by SP6 and T7 RNA polymerases from specific DNA templates is described. Transcription of templates prepared by digestion with restriction enzymes that leave 3' protruding ends resulted in the production of significant
D A Melton et al.
Nucleic acids research, 12(18), 7035-7056 (1984-09-25)
A simple and efficient method for synthesizing pure single stranded RNAs of virtually any structure is described. This in vitro transcription system is based on the unusually specific RNA synthesis by bacteriophage SP6 RNA polymerase which initiates transcription exclusively at
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