Vaccinia Capping Enzyme
Cat.No:R8610 Solarbio
Storage:-20℃
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Storage | -20℃ |
Background | Capping buffer: 0.5 M Tris-HCl, 50 mM KCl, 10 mM MgCl2, 10 mM DTT, (25℃, pH8.0). |
Note | 1. The efficiency of this capping reaction is affected by the structure of the RNA 5' end, it is recommended to open the advanced structure of the 5' end of RNA by thermal denaturation (heating at 65℃ for 5 min, placing on ice for 5 min). 2. This capping reaction can be completed within 1hour generally. If the structure of the RNA 5' end is complex or the RNA length is short (≤ 200 nt), the reaction time can be extended to 2hours. 3. SAM is a high-energy methyl donor, which is unstable in nature. It is recommended to thaw and add samples on ice in the whole process. 4. If more RNA is capped with the same amount of enzyme, the incubation time at 37℃ can be appropriately extended to 2hours. |
Source | Recombinant E.coli |
Unit Definition | 1U VCE refers to the amount of enzyme required to incorporate 10pmol GTP into RNA for 1 hour at 37℃ |
Concentration | 10U/ul |
Solvent | 20 mM Tris-HCl pH 8.0,100 mM NaCl,1 mM DTT,0.1 mM EDTA,0.1%(v/v)Triton X-100,50%(v/v)Glycerol |
Unit | Piece |
Specification | 500U 2KU |
Vaccinia Capping Enzyme, derived from recombinant E.coli strain carrying vaccinia virus MR, consists of two subunits D1R and D12L with three enzyme activities (RNA triphosphatase activity, guanylate transferase activity and guanine methyltransferase activity). Vaccinia Capping Enzyme can effectively catalyze the formation of the cap structure, which can specifically add the 7-methylguanosine cap structure (m7Gppp, Cap 0) to the 5' end of RNA. Cap0 plays an important role in the stability, transport and translation of eukaryotic mRNA. Capping RNA by enzymatic reactions is an effective and simple method that can significantly improve the stability and translation ability of RNA for in vitro transcription, transfection, and microinjection.
Remark:These protocols are for reference only. Solarbio does not independently validate these methods.
Note:
1. The products are all for scientific research use only. Do not use it for medical, clinical diagnosis or treatment, food and cosmetics, etc. Do not store them in ordinary residential areas.
2. For your safety and health, please wear laboratory clothes, disposable gloves and masks.
3. The experimental results may be affected by many factors, after-sale service is limited to the product itself and does not involve other compensation.
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