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T7 RNA Polymerase, CAS No.9014-24-8

COA COA
  • Accession #: P00573
In stock
Item Number
T292917
Grouped product items
SKU Size
Availability
 Price Qty
T292917-50μl
50μl
Available within 8-12 weeks(?)
Production requires sourcing of materials. We appreciate your patience and understanding.
$39.90
T292917-250μl
250μl
Available within 8-12 weeks(?)
Production requires sourcing of materials. We appreciate your patience and understanding.
$159.90
T292917-1ml
1ml
Available within 8-12 weeks(?)
Production requires sourcing of materials. We appreciate your patience and understanding.
$539.90
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View related series
Click to view protein information: POLRMT
Cell Cycle (2830) RNA Polymerase (1)

Basic Description

Product Name T7 RNA Polymerase, CAS No.9014-24-8
Synonyms T7 RNA Polymerase
Grade BioReagent, for molecular biology, RNase free, EnzymoPure™
Product Description

T7RNA polymerase is a DNA-dependent RNA polymerase derived from T7 phage with highly specific 5'→3' terminal RNA polymerase activity. T7RNA polymerase has high specificity to the T7 promoter and can synthesize a large amount of RNA using the downstream sequence of the T7 promoter as a template.
Product Content

T292917Component50 μL250 μL1 mLStorage
T292917AT7 RNA Polymerase50 μL250 μL1 mL-20°C. Avoid freeze/thaw cycle
T292917B10×HH T7 Buffer2×50 μL2×250 μL2×1 mL-20°C. Avoid freeze/thaw cycle

Activity unit definition
In a standard reaction system, the amount of enzyme required to catalyze NTP to produce a 1nmol PPi within 37°C1 hours is defined as one unit of activity (U).
Iinstructions
System configuration (50 μ l）

组分体积
Nuclease-free water/DEPC
Up to 20 μL
10×T7 RNA Polymerase Buffer
2 μL
ATP/GTP/CTP/UTP (100mM each)
0.4 μL each (2mM each Final)
RNase inhibitor
1 μL (40 U)
Inorganic pyrophosphatase
0.5 μL (0.05 U)
T7 RNA Polymerase (50 U/μL)
1 μL
Linearized template DNA
1 μg

10×HH T7 Buffer: 400 mM Tris-HCl (25℃，pH 8.20), 60 mM MgCl2, 100mM DTT, 20mM spermidine.

The addition of reaction components *10xHH T7 Buffer in the above order is only applicable to the dosage of NTPS with a final concentration of 2mM. If the dosage of 7.5mM to 10mM is required, please choose the high-yield kit series. 

Reaction time: Incubation at 37℃ for 2 h.
Reaction termination: Add 2μL 0.2M EDTA (pH = 8.0@25 °C) or heat at 75° C for 10 min. 

DNA template removal: DNasel(2 U) at 37°C for 15 min.
Quality Control
1) Endonucrenase residue detection: 50U benzyme and 1μg λDNA were added to the 50μL reaction system at 37°C for 16 hours, and the DNA bands of agarose gel electrophoresis did not change.
2) Exonuclide residue detection: 50U benzyme and 1μgλ-HindIIIdigestDNA were added to the 50μL reaction system and incubated at 37°C for 16 hours, and the DNA bands of agarose gel electrophoresis did not change.
3) Nucleic acid incision enzyme residue detection: 50U benzyme and 1μgpBR322DNA were added to the 50μL reaction system and incubated at 37°C for 16 hours, and the DNA bands of agarose gel electrophoresis did not change.
4) RNase residue detection: 50U benzyme and 1.6μgMS2RNA were added to the 50μL reaction system and incubated at 37°C for 16 hours, and the RNA bands of agarose gel electrophoresis did not change.
5) Thermal inactivation: 75°C, 10min.

Matters needing attention
1. The preparation of transcription reaction should be completed in an environment free of RNase pollution, and gloves are recommended during operation. The system was prepared using nuclease-free water, suction heads and reaction tubes.
2, you can increase the RNA production by increasing the NTP concentration (each concentration can reach 10mM), while the need to appropriately increase the Mg "concentration.
3, the reaction system needs to be prepared at room temperature to avoid DNA reaction with 10xHH T7 Buffer at 4 ℃ and precipitate.
4. Incomplete linearization of template DNA may reduce the yield and length of transcription products.
5, the volume of the reaction system can be scaled up or down according to actual needs.
6. If the reaction product decreases, 20 mM fresh DTT can be added to the reaction system.
7, transcription fragments ≤500bp, it is recommended to extend the transcription time to 4-8 h.
8. 10xHH T7 Buffer is prone to precipitate after freezing. Please confirm whether it is completely dissolved before use. If precipitate occurs, it can be shaken and redissolved after fully heating at 37 ℃
9, 10xHH T7 Buffer is only suitable for 2mM final concentration of NTPS feed, such as 7.5mM-10mM feed, please choose high yield kit series products.
Specifications & Purity BioReagent, EnzymoPure™, RNase free, Suitable for molecular biology, ≥98.0%, 50 U/μL
Accession # P00573

Storage and Shipping

Concentration 50 U/μL
Storage Temp Store at -20°C,Avoid repeated freezing and thawing
Shipped In Ice chest + Ice pads
Stability And Storage Store at -20℃ long term (24 months). Upon receipt, it is recommended to aliquot. Avoid freeze/thaw cycle.
CAS 9014-24-8
Enzyme Commission Number 2.7.7.6
Unit definition In a standard reaction system, the amount of enzyme required to catalyze NTP to produce a 1nmol PPi within 37°C1 hours is defined as one unit of activity (U).

Certificates（CoA,COO,BSE/TSE and Analysis Chart)

C of A & Other Certificates(BSE/TSE, COO):
Analytical Chart:

Alternative Products

  1. Thermostable T7 RNA Polymerase
    Thermostable T7 RNA Polymerase

    Starting at $447.90

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