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mRNA Cap 2´-O-Methyltransferase, GMP Grade(Cat. No.:GMP-M072)
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Description
  • With S-adenosylmethionine (SAM) as the methyl group base donor, mRNA Cap 2´-O-Methyltransferase can add a methyl group to the 2´-O of the first nucleotide of the Cap0 structure, [m7GpppN1(pN)x-OH(3')], next to the RNA 5´ end. Methylation of capped mRNA from Cap 0 to Cap1 naturally exists in eukaryocyte, where the Cap1 structure shows an important effect for translation, thus for expression as well. This product doesn’t show a reactivity to RNA substrate other than RNA with Cap0 structure.
  • This product is from a large scale GMP leveled recombinant Cap 2’-O-Methyltransferase production via E. coli expression. Applying pharmaceutical leveled adjuvant and material for production, strictly controlling host protein residues, nucleic acid residues and other impurities, we guarantee manufacture and quality control practice complying to GMP regulation, as well as all the materials traceable.
  • This product has completed the DMF record of FDA and passed the HALAL certification.
Usage
Methylate the 2´-O of the first nucleotide of the Cap0 structure, to realize mRNA translation and expression.
Quality Control
ElementStandard
AppearanceClear and transparent solution
IdentificationPositive
Visible ParticlesMeet the specification
pH7.5-8.5
Activity50KU/ml-52.5KU/ml
Purity≥95%
Protein ContentMeet the specification
Endonuclease residuesThe degradation of substrate was ≤10%
Exonuclease residuesThe degradation of substrate was ≤10%
RNase ResiduesThe degradation of substrate was ≤10%
Bacterial Endotoxins<5EU/ml
Exogenous DNA Residues≤100pg/mg
Host-cell Protein Residues≤50ppm
MycoplasmaNegative
Heavy Metals≤10ppm
Microbial LimitTotal aerobic microbial count≤1cfu/10ml, total yeasts and molds count≤1cfu/10ml
Storage
At -20±5℃.
Components
SKUComponentsVolume
GMP-M072-01AmRNA Cap 2’-O-Methyltransferase, GMP Grade (50U/μl)50μl
GMP-M072-M001mRNA Cap 2’-O-Methyltransferase, GMP Grade (50U/μl)1ml
GMP-M072-M010mRNA Cap 2’-O-Methyltransferase, GMP Grade (50U/μl)10ml
GMP-M072-M050mRNA Cap 2’-O-Methyltransferase, GMP Grade (50U/μl)50ml
Notes
  • SAM is unstable at pH 7–8, 37 °C and needs to be freshly prepared before the reaction starts. The amount of SAM can be calculated in advance, and the aliquoted 32 mM stock solution is diluted into a 2mM working solution before the reaction starts. To avoid SAM degradation, the working solution needs to be kept on ice.
  • Prepare Cap0-capped RNA for Cap0 to Cap1 methylation. Before starting reaction, the RNA applied for capping should be purified and dissolved in nuclease-free water before. The solution for use must NOT contain any EDTA or salt. We may obtain Cap0-capped RNA by Vaccinia Capping System (GMP-M062, Novoprotein). We may run both Cap0 capping and Cap1 capping in one reactor at the same time. Equally, we may realize cap1 capping conveniently by applying capping reaction kit (M082, Novoprotein).
  • RNA transcript with 5’ end secondary structure can be an obstacle for capping. Heating the RNA sample can remove complex structure of the transcript. We may extend heating time to 10 minutes, and extend the capping reaction time to 60 minutes.
  • When used for the capped RNA 3’-poly (A) tailing, we may apply E. coli Poly(A) Polymerase, GMP Grade (GMP-M012, Novoprotein).
  • The sample needs to be purified before transfection.
  • When adding 0.5 µl of RNase inhibitor (GMP-E125, Novoprotein) to the reaction system (shown below in “Manual” part), please deduct equal volume of RNase-free water.
  • Do NOT store the product at -70°C or an even lower temperature.
  • In case of white precipitation in reaction buffer (shown below in “Manual” part), incubate it at 37 ° C for 5 mins, blend it, totally dissolving precipitation.
Related Products
Cat. No.Product NameCat. No.Product Name
GMP-M062Vaccinia Capping Enzyme, GMP GradeGMP-E125RNase Inhibitor, GMP Grade
GMP-M036Pyrophosphatase, Inorganic (yeast), GMP GradeGMP-E127DNase I, GMP Grade
GMP-M012E. coli Poly(A) Polymerase, GMP GradeGMP-E121T7 RNA Polymerase, GMP Grade
GMP-EB6210x Capping Reaction Buffer, GMP GradeGMP-S062SAM, GMP Grade
GMP-S024NGTP, GMP Grade
仅供科研或生产使用,不可直接应用于人体。
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