This product contains vaccinia virus capping enzyme, mRNA cap structure 2'-O-methyltransferase and other capping reaction components, which can be used for 5' capping modification of RNA produced by T7 in vitro transcription (JT101).
In eukaryotes, after transcription to form the original mRNA, a special cap structure needs to be formed at the 5' end. This structure plays an important role in the stability, transport (exit) and translation of mRNA. The capping modification of the 5' end of RNA by the enzymatic reaction of capping enzymes is a simple and effective method: Vaccinia Capping Enzyme can attach 7-methylguanylate cap (m7Gppp, Cap0) to the 5' end of RNA to form m7Gppp5'N-mRNA (Cap0-mRNA). The mRNA Cap 2 '-O-methyltransferas uses CAP0-mRNA as substrate and SAM (S-adenosine methionine) as a methyl donor to methylate 2' -oh of the first nucleotide of cap0-mrna 5 'end adjacent to the cap structure to form Cap1-mRNA.
Cap1 structure can improve mRNA stability, which helps to enhance its expression ability in cell transfection and microinjection experiments.
This product uses vaccinia virus capping enzyme and mRNA Cap-2'-O-methyltransferase at the same time in the capping reaction, so that the capping reaction can be completed in the same reaction. The reaction product is Cap1-mRNA, and the capping efficiency can be close to 100% in the correct cap orientation.
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