Faustovirus capping enzyme

FCE
Catalog No. ON-981-C025
Storage Condition -15 ~ -25℃
Source Recombinant E.coli
Purity >95% by SDS-PAGE

Faustovirus capping enzyme

FCE
Grade
SKU: ON-981-C025
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Details

Product Information

Product description: Faustavirus capping enzyme (FCE) is encoded by Faustavirus. FCE is a single-subunit enzyme that possesses the three enzymatic activities required for capping RNA with a Cap-0 structure: triphosphatase, guanylyltransferase, and (guanine-N7)-methyltransferase. FCE has a wider reaction temperature range compared to vaccinia capping enzyme (VCE). It can perform capping at temperatures ranging from 37°C to 50°C. FCE acts synergistically with mRNA Cap 2'-O-Methyltransferase to add a Cap-1 structure to mRNA in a single step.

Source: An E. coli strain that carries FCE gene.

Concentration: 25 units/µL

Unit Definition: One unit is defined as the amount of enzyme that converts 25pmols of a 80nt transcript to Cap-0 RNA in 30minutes at 37°C.

Storage Buffer: 40mM Tris-HCl, 100mM NaCl, 50mM Arginine, 0.1mM TCEP, 50% Glycerol, (pH8.0@25°C).

Companion Product: 10X FCE Reaction Buffer: 500mM Tris-HCl, 50mM KCl, 10mM MgCl2, 10mM DTT, pH8.0@25℃.

Key points of operation

1. Do not oscillate vigorously when mixing.

2. If the reagents, tubes, or micropipette tips are contaminated with RNase, it will result in reduced RNA yield or degradation. Precautions should be taken to avoid RNase contamination during experiments, such as wearing disposable gloves and using special pipettes.

3. The mRNA solution was heated at 65ºC for 5 minutes to open the secondary structure at the 5' end of the transcription product before warming with FCE. For transcription products with highly structured 5' ends, the time can be extended to 10 minutes or the denaturation temperature can be increased appropriately.

4. SAM is unstable at pH 7-8 and 37ºC. It is recommended to prepare fresh working solution according to the actual number of reactions and place it on ice before the start of the reaction to prevent SAM degradation.

Security Information

Storage Conditions: -15 ~ -25℃

Quality Assurance: Free of Endonuclease, Exonuclease and RNase activities.

Physical Purity: >95% by SDS-PAGE.


Reference

1. Ramanathan, A. et al. (2016). Nucleic Acids Res . 44 (16), 7511–7526.

2. Furuichi, Y., LaFiandra, A. & Shatkin, A. (1977). Nature. 266, 235-239.

Specifications

  • Catalog No.
    ON-981-C025
  • Source
    Recombinant E.coli
  • Purity
    >95% by SDS-PAGE
  • Storage Condition
    -15 ~ -25℃

Documentation

Product Information

Product description: Faustavirus capping enzyme (FCE) is encoded by Faustavirus. FCE is a single-subunit enzyme that possesses the three enzymatic activities required for capping RNA with a Cap-0 structure: triphosphatase, guanylyltransferase, and (guanine-N7)-methyltransferase. FCE has a wider reaction temperature range compared to vaccinia capping enzyme (VCE). It can perform capping at temperatures ranging from 37°C to 50°C. FCE acts synergistically with mRNA Cap 2'-O-Methyltransferase to add a Cap-1 structure to mRNA in a single step.

Source: An E. coli strain that carries FCE gene.

Concentration: 25 units/µL

Unit Definition: One unit is defined as the amount of enzyme that converts 25pmols of a 80nt transcript to Cap-0 RNA in 30minutes at 37°C.

Storage Buffer: 40mM Tris-HCl, 100mM NaCl, 50mM Arginine, 0.1mM TCEP, 50% Glycerol, (pH8.0@25°C).

Companion Product: 10X FCE Reaction Buffer: 500mM Tris-HCl, 50mM KCl, 10mM MgCl2, 10mM DTT, pH8.0@25℃.

Key points of operation

1. Do not oscillate vigorously when mixing.

2. If the reagents, tubes, or micropipette tips are contaminated with RNase, it will result in reduced RNA yield or degradation. Precautions should be taken to avoid RNase contamination during experiments, such as wearing disposable gloves and using special pipettes.

3. The mRNA solution was heated at 65ºC for 5 minutes to open the secondary structure at the 5' end of the transcription product before warming with FCE. For transcription products with highly structured 5' ends, the time can be extended to 10 minutes or the denaturation temperature can be increased appropriately.

4. SAM is unstable at pH 7-8 and 37ºC. It is recommended to prepare fresh working solution according to the actual number of reactions and place it on ice before the start of the reaction to prevent SAM degradation.

Security Information

Storage Conditions: -15 ~ -25℃

Quality Assurance: Free of Endonuclease, Exonuclease and RNase activities.

Physical Purity: >95% by SDS-PAGE.


Reference

1. Ramanathan, A. et al. (2016). Nucleic Acids Res . 44 (16), 7511–7526.

2. Furuichi, Y., LaFiandra, A. & Shatkin, A. (1977). Nature. 266, 235-239.

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