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    CleanT7™ RNA polymerase (large pack)

    CleanT7™ RNA polymerase (large pack)

    CleanT7™ RNA polymerase is an engineered variant of T7 RNA polymerase with significant reduction in double-stranded RNA (dsRNA) by-product. CleanT7 shares other biochemical properties with wild-type T7 RNA polymerase, such as promoter sequence, IVT reaction condition and compatibility with CAP analogs.

    Highlights:

    1. Highly pure in vitro transcribed RNA, with much reduced dsRNA as compared to wild-type T7 RNA polymerase
    2. Easy migration by only swapping in the CleanT7 RNA polymerase itself

    Application:

    1. In vitro synthesis of highly pure RNA of all kinds

    Product information

    Product source:

    Purified from E.coli carrying a plasmid which contains the CleanT7 RNA Polymerase gene. 

    Contents provided:

     item concentration quantity
    CleanT7™ RNA polymerase 50U/ul

    2500U(RT002S)

    12500U(RT002L)
    10x reaction buffer 10x

    150ul(RT002S)

    600ul(RT002L)

    Instruction for use

    CleanT7 RNA polymerase Instruction for Use

    FAQs

    1. Can CleanT7™ RNA polymerase incorporate modified NTP?

    Similar to wild-type T7 RNAP, CleanT7™ RNA polymerase can efficiently incorporate modified NTP, such as 5-methylcytidine (5mC), 5-methoxyuridine (5moU), and pseudouridine (Ψ).

    2. Can CleanT7™ RNA polymerase incorporate CleanCap reagents?

    CleanT7™ RNA polymerase supports co-transcriptional capping using CleanCap(TriLink Biotechnologies).

    3. How to detect dsRNA in IVT product?

    Here is an example protocol:

    1. RNA transcripts (200 ng) were dropped onto Immobilon™ -Ny+ Membrane (Millipore), dried, blocked with 5% non-fat dried milk in TBS-T buffer (50 mM Tris–HCl pH 7.4, 150 mM NaCl, 0.05% Tween-20).
    2. Incubated with dsRNA-specific mAb J2 (English & Scientific Consulting) for 30 min at 25°C. Membranes were washed 2 times with TBS-T buffer and reacted with HRP-conjugated donkey anti-mouse Ig (Jackson Immunology), washed 2 times.
    3. Add the ECL Western blot detection reagent (Thermo) onto the Ny+ Membrane. fluorescence signals were captured by X film (Kodak) photosensitive development or other fluorescence imager instrument

     

     

    Reference

    Bingbing Yu, Yifan Chen, Yan Yan, Xueling Lu and Bin Zhu DNA-t erminus-dependent tr anscription b y T7 RNA polymerase and its C-helix mutants. Nucleic Acids Research, 2024 (link)

    Legal and disclaimers

    This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by founders of RiboTech.

    This product is intended for research use only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

    Shipping

    Products are shipped with regular gel ice pack in an insulated foam box. We ship via next day air service (Monday to Thursday).

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