商品编号


M0366S-2,000 units



品牌


纽英伦(NEB)



公司


New England Biolabs



公司分类


RNA Reagents



Concentration

50,000 units/ml


Size

2,000 units


商品信息

Description:
mRNA Cap 2?-O-Methyltransferase adds a methyl group at the 2?-O position of the first nucleotide adjacent to the cap structure at the 5? end of the RNA. The enzyme utilizes S-adenosylmethionine (SAM) as a methyl donor to methylate capped RNA (cap-0) resulting in a cap-1 structure.
The cap-1 structure has been reported to enhance mRNA translation efficiency (1) and hence may help improve expression in mRNA transfection and microinjection experiments.
mRNA Cap 2?-O-Methyltransferase specifically requires RNA with an m7GpppN cap as substrate. It cannot utilize RNA with pN, ppN, pppN or GpppN at the 5? end (2,3). Capped RNA may be prepared via in vitro transcription using cap analog or through enzymatic capping using the Vaccinia Capping Enzyme (
NEB
#M2080 ).
The reagents provided in this pack can be used to methylate up to 400 μg of capped RNA.
?
?
Figure 1. 5? Cap Structure
Schematic representation of mRNA 5? cap structure indicating the 7-methylguanosine, shown in yellow, and the 5? end of the mRNA, shown in blue. The 2?-O-methyl group present in Cap 1 structures is shown in red.
Product Source
An
E. coli
strain that carries the gene for the Vaccinia mRNA Cap 2?-O-Methyltransferase.
Reagents Supplied
The following reagents are supplied with this product:
Store at (°C)
Concentration
S-adenosylmethionine (SAM)
-20
32 mM
Capping Buffer
10X
Notes:
(READ PRIOR TO SETTING UP REACTION)RNA prepared using in vitro transcription and cap analog should be purified prior to use and res
USP
ended in nuclease-free water. EDTA and salts should not be present in the solution. mRNA Cap 2?-O-Methyltransferase may be directly added to a Vaccinia Capping System (
NEB
#M2080) reaction. RNA purification is not required in this case.Heating the RNA at 65°C for 5 minutes prior to incubation with the enzyme removes secondary structure on the 5? end of the transcript. Extend time to 10 minutes for transcripts with known highly structured 5? ends.SAM is unstable at pH 7–8, 37°C and should be mixed fresh prior to starting the reaction. We recommend determining how many reactions will be performed and diluting an aliquot of the 32 mM stock to 4 mM immediately before setting up the reactions. This "working stock" should be kept on ice to prevent degradation of SAM.