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mRNA Cap 2´-O-Methyltransferase GMP-grade (50 U/μl)
产品货号:10612ES92
产品规格:10,000 U
产品价格:¥ 4285
类别:体外转录系列

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  • Product Information

     

    Product Name

    Catalog No.

    Size

    Price ()

    mRNA Cap 2´-O-Methyltransferase GMP-grade (50 U/μL)

    10612ES92

    10,000 U

    4285

    10612ES97

    50,000 U

    18585

    10612ES98

    250,000 U

    Inquiry

    10612ES99

    20 MU

    Inquiry

     

    Product Description

     

    mRNA Cap 2´-O-Methyltransferase is a recombinant protein derived from vaccinia virus. The enzyme can add a methyl group at the 2´-O position of the first nucleotide of the cap structure at the 5´ end of the RNA. The enzyme uses SAM as a methyl donor to methylate capped RNA to form a Cap1 structure. The Cap1 structure can enhance the translation efficiency of mRNA, so it can improve the expression of mRNA in transfection and microinjection experiments. This enzyme requires RNA with m7GpppN, a 7-methylguanosine cap structure, as a substrate.

    This product is produced in accordance with GMP process requirements. The product is provided in liquid form and can be used for Cap1 capping reaction of pre-mRNA in vivo/in vitro.

     

    Product Properties

     

    Source

    Recombinant E.coli with Cap 2´-O-Methyltransferase gene

    Optimum Temperature

    37℃

    Storage Buffer

    20 mM Tris-HCl pH8.00.1mM EDTA1mM DTT100mM NaCl50%v/vglycerin0.1%v/vTrion X-100  

    Unit Definition

    One unit is defined as the amount of enzyme required to methylate 10 pmol of 80nt capped RNA transcript in 1 h at 37°C.

     

    Contents

     

    Contents No.

    Name

    Catalog No./Specification

    10612ES92

    (10,000 U)

    10612ES97

    (50,000 U)

    10612ES98

    (250,000 U)

    10612ES99

    (20 MU)

    10612

    mRNA Cap 2´-O-Methyltransferase GMP-grade (50 U/μL)

    200 μL

    1 mL

    5 mL

    400 mL

     

    Shipping and Storage

     

    The mRNA Cap 2´-O-Methyltransferase GMP-grade products are shipped with dry ice and can be stored at -15℃ ~ -25℃ for one year.

     

    Experimental methods

     

    Cap1 capping reaction (20 μL reaction system)

    This step is suitable for capping reaction of 10 μg RNA (≥ 100 nt), and can be amplified according to experimental needs.

    1) Take 10 μg RNA to a 1.5 mL centrifuge tube and dilute to 9.5 μL with nuclease-free water;

    2) Heat at 65°C for 5 min;

    3) Take out the centrifuge tube and place it on ice for 5 min;

    4) Add the following components in sequence:

    Components

    Volume

    Denatured RNA

    9.5 μL

    10×Capping Buffer

    2.0 μL

    Murine RNase inhibitor(40 U/μL)

    0.5 μL

    GTP (10 mM)

    1.0 μL

    SAM (10 mM, fresh)

    1.0 μL

    Vaccinia Capping Enzyme (10 U/μL)

    5.0 μL

    Cap 2´-O-Methyltransferase (50 U/μL)

    1.0 μL

    Note10×Capping Buffer(Cat# 10666)0.5 M Tris-HCl, 50 mM KCl, 10 mM MgCl2, 10 mM DTT pH 8.0 @ 25℃.

    5) Incubate at 37°C for 2 h;

    6) RNA capping is completed, next experiments can be performed.

     

    Notes

     

    1. For your safety and health, please wear personal protective equipment (PPE), such as laboratory coats and disposable gloves, when operating with this product.

    2. The extracted RNA needs to be purified and resuspended in nuclease-free water;

    3. The RNA solution needs to be heated before adding the enzyme to remove the secondary structure at the 5'end;

    4. For RNA with a known 5'end structure, the reaction time can be extended to 4 h to improve the capping efficiency;

    5. In the 5'end labeling reaction system, the GTP stock solution should be diluted to 1-3 times of the mRNA molar concentration in the reaction system.

     

     

    HB220514