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    Ice lake™ (VSW-3) RNA polymerase (large pack)

    Ice lake™ (VSW-3) RNA polymerase (large pack)

    Ice lake™(VSW-3) RNA polymerase is a DNA-dependent RNA polymerase. It is isolated from a psychrophilic phage VSW-3. Ice lake RNA polymerase is capable of in vitro synthesis of RNA at low temperature (4-25℃). Compared with wild-type T7 RNA polymerase, Ice lake™ RNA polymerase is an ideal alternative for producing RNA with higher purity.

    Promoter sequence: TTAATTGGGCCACCTATA

    Comparison with T7 promoter:

    Highlights:

    1. Highly pure in vitro transcribed RNA, less by-products such as full-length or short double-stranded RNA and 3'-addition compared to T7 RNA polymerase

    2. Low temperature in vitro transcription to maintain stability of RNA

    Application:

    1. In vitro synthesis of highly pure mRNA
    2. in vitro synthesis of modified RNA

     

    Product information

    Product source:

    Purified from E.coli carrying a plasmid which contains the VSW-3 RNA Polymerase gene. 

    Contents provided:

     item concentration quantity
    Ice Lake™(VSW-3) RNA polymerase 50U/ul

    2500U(RT001S)

    12500U(RT001L)
    5x reaction buffer 5x

    150ul(RT001S)

    600ul(RT001L)

    Instruction for use

    VSW-3 Manual

    FAQs

    1. Can Ice lake™ RNA polymerase incorporate modified NTP?

    Ice lake™ RNA polymerase can efficiently incorporate modified NTP, such as 5-methylcytidine (5mC), 5-methoxyuridine (5moU), and pseudouridine (Ψ).

    2. Can Ice lake™ RNA polymerase incorporate CleanCap reagents?

    Currently Ice lake™ RNA polymerase does not support co-transcriptional capping using CleanCap(TriLink Biotechnologies). Capping of in vitro transcribed RNA should be done by using capping enzymes per manufacturer's instructions.

    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

    Here is an example result:

     

     

    Reference

    1. Heng Xia, Bingbing Yu, Yixin Jiang, Rui Cheng, Xueling Lu, Hui Wu, Bin Zhu Psychrophilic phage VSW-3 RNA polymerase reduces both terminal and full-length dsRNA byproducts in in vitro transcription. RNA Biology, 2022, 19(1), 1130–1142 [link]
    2. Guoquan Wang, Rui Cheng, Qiubing Chen, Yuandong Xu, Bingbing Yu, Bin Zhu, Hao Yin, Heng Xia mRNA produced by VSW-3 RNAP has high-level translation efficiency with low inflammatory stimulation. Cell Insight, Volume 1, Issue 5, October 2022 [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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