Product Class: Kit

HiScribe™ T7 ARCA mRNA Kit
Item# Description List Price Web Price Qty
E2065S HiScribe™ T7 ARCA mRNA Kit $447.00
$402.30
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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S1407S G(5')ppp(5') RNA Cap Structure Analog - 1 umol $169.00
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S1411L 3'-O-Me-m7G(5')ppp(5')G RNA Cap Structure Analog - 5 umol $841.00
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    Product Introduction

    • Generate up to 20 μg of capped mRNA per reaction
    • mRNA capping, DNA removal, mRNA tailing and purification complete in 2 hrs. Competitive products have more pipetting steps and separate components.
    • Enables partial incorporation of 5mCTP, Pseudo-UTP and other modified CTP and UTP
    • Ultra-high quality components ensure mRNA integrity
    • ARCA-based capping in correct orientation ensures high translation efficiency
    • Template removal and mRNA purification reagents included
    • HiScribe™ kits contain twice the number of reactions as competitive products
    • Also available with tailing reagents

    Product Information

    Description

    Most eukaryotic mRNAs require a 7-methyl guanosine (m7G) cap structure at the 5´ end and a Poly(A) tail at the 3´ end to be efficiently translated. By using a DNA template encoding a poly(A) tail, the HiScribe T7 ARCA mRNA Kit can be used to synthesize capped and tailed mRNAs. The cap structure is added to the mRNA by co-transcriptional incorporation of Anti-Reverse Cap Analog (ARCA) (NEB #S1411) using T7 RNA Polymerase. Poly(A) tail is incorporated during the transcription reaction. The kit also includes DNase I and LiCl for DNA template removal and quick mRNA purification.

    Additionally, the kit is capable of partial incorporation of modified UTP and CTP (up to 50% each) without affecting the mRNA yield significantly. By using a DNA template encoding a poly(A) tail, capped and tailed modified mRNA can be synthesized in a single reaction in 30 minutes. mRNAs synthesized with the kit can be used for cell transfection, microinjection, in vitro translation and RNA vaccines.

    ARCA is incorporated into mRNA exclusively in the correct orientation, generating capped mRNA that is more efficiently translated. Standard cap analogs can be incorporated in either direction resulting in only 50% of capped mRNA that is functional in protein translation.

    Figure 1. Structure of Anti-Reverse Cap Analog (ARCA, NEB #S1411)

    Methylation at the 3´ position of 7mG forces the cap structure to be attached to mRNA in the correct orientation.
    Figure 2. Overview of mRNA synthesis work flow with the HiScribe T7 ARCA mRNA KitFigure 2. Overview of mRNA synthesis workflow with the HiScribe T7-ARCA mRNA Kit

    Kit Components

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    LiCl Solution -20
    DNase I (RNase-free) -20 2 units/µl
    T7 RNA Polymerase Mix -20
    CLuc Control Template -20 0.25 µg/µl
    ARCA/NTP Mix -20 2X
    Product Categories:
    RNA Synthesis Products
    Applications:
    RNA Modification,
    In vitro Synthesis (IVT)

    Properties & Usage

    Materials Required but not Supplied

    • DNA template
    • Thermocycler or 37°C incubator.
    • Nuclease-free water
    • Buffer- or water-saturated phenol:chloroform
    • Ethanol
    • 3 M Sodium acetate, pH 5.2
    • 5 M Ammonium acetate
    • Spin columns
    • Gels, running buffers and gel box
    • Equipment for RNA analysis

    Storage Temperature

    -20°C

    Protocols, Manuals & Usage

    Protocols

    1. Standard mRNA Synthesis (E2065)
    2. mRNA Synthesis with Modified Nucleotides (E2065)
    3. mRNA Purification (E2065)
    4. Evaluation of Reaction Products (E2065)

    Manuals

    The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.

    Faqs & Troubleshooting

    FAQs

    1. HiScribe™ T7 ARCA mRNA Kit (with tailing) What is the difference between the HiScribe T7 ARCA mRNA Kit (NEB #E2065) and the HiScribe T7 ARCA mRNA Kit (with Tailing)(NEB #E2060)?
    2. I currently use MessageMAX™ T7 ARCA-capped Message Transcription Kit, which mRNA synthesis kit from NEB should I use?
    3. I currently use mMessage mMachine® T7 Ultra Transcription Kit, which mRNA synthesis kit from NEB should I use?
    4. Can modified nucleotides be used with the HiScribe T7 ARCA mRNA kits?
    5. What is the difference between the HiScribe T7 ARCA mRNA kits and the HiScribe T7 High Yield RNA Synthesis Kit (E2040) and HiScribe T7 Quick RNA Synthesis Kit (E2050)?
    6. Can I use the Monarch RNA Cleanup Kits to cleanup my in vitro transcription (IVT) reaction?

    Troubleshooting

    Quality, Safety & Legal

    Quality Assurance Statement

    Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

    Specifications

    The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

    Certificate Of Analysis

    The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

    Safety DataSheets

    The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

    Legal and Disclaimers

    This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

    While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

    For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

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