Active Recombinant 2'-O-Methyltransferase

• Cat.No.: 2'-O-Methyltransferase-01
• Product Overview: 2'-O-Methyltransferase was expressed in E.coli. DNA Exonuclease Activity: Negative DNA Endonuclease Activity: Negative RNase Activity: Negative Residual Host Cell DNA: <100 pg/mg

Specification

• Source: E.coli
• Tag: Non
• Description: 2'-O-Methyltransferase can utilize S-adenosylmethionine (SAM) as a methyl donor to add a methyl group to the 2'-O position of the first nucleotide adjacent to the cap structure (Cap0) at the 5'-end of RNA, forming the Cap1 structure. It can only use RNA with a 7-methylguanosine cap structure (m7GpppN) as a substrate and will not act on RNA with pN, ppN, pppN, or GpppN at the 5'-end. The Cap1 structure can enhance the translation efficiency of mRNA and contribute to improving the expression level during the mRNA transfection process. It is a necessary modification for in vitro mRNA synthesis.
• Purity: ≥ 95% by SDS-PAGE
• Bio-activity: ≥ 100 U/μL
• Advantages: 1. It is capable of capping RNA within one hour, and the capping efficiency can reach 100%. 2. There is no contamination of exogenous DNase and RNase.
• Unit definition: 1 U is defined as the amount of enzyme required to methylate 10 pmol of an 80 nt capped RNA transcription product within 1 hour at 37 centigrade.
• Usage: 1. Dilute the Cap0 RNA to 16 μL (up to 10 μg) with RNase-free water. 2. Heat at 65 centigrade for 5 minutes. 3. Place on ice for 5 minutes. 4. Prepare the following reaction system. Cap0 RNA: 16.0 μL 10× 2'-O-Methyltransferase reaction buffer: 2.0 μL SAM (2 mM, pre-diluted from the 32 mM stock solution): 1.0 μL 2'-O-Methyltransferase: 0.5-1.0 μL Total volume: 20.0 μL 5. Incubate at 37 centigrade for 60 minutes. For RNAs shorter than 200 nt, it is recommended to extend the reaction time to 120 minutes. It is advisable to add RNase Inhibitor during the reaction process.
• Applications: The 5'-end capping reaction of mRNA in vivo/in vitro (conversion from Cap0 to Cap1)
• Note: 1. Try to avoid freeze-thaw cycles of this product after receipt; 2. Please wear lab coat and disposable gloves when using; 3. This product should not be used directly for clinical diagnosis and treatment.
• Stability: This product should be stored at -20 centigrade and can be stored for at least 24 months.
• Storage: Store at -20 centigrade.
• Storage Buffer: 20 mM Tris-HCl, 100 mM NaCl, 1 mM DTT, 0.1 mM EDTA, 0.1% TritonX-100, 50% Glycerol, pH8.0
• Shipping: Dry Ice

Gene Information

• Official Symbol: 2'-O-Methyltransferase
• Synonyms: 2'-O-Methyltransferase