HiScribe™ T7 ARCA mRNA Kit (with tailing)#E2065

Catalog # Size Price (£)
E2060S 20 reactions £342.00
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. The HiScribe T7 ARCA mRNA Kit (with tailing) is designed for quick production of ARCA capped and poly(A) tailed mRNA in vitro. Capped mRNAs are synthesized by co-transcriptional incorporation of Anti-Reverse Cap Analog (ARCA, NEB #S1411) using T7 RNA Polymerase. The transcription reaction can be set up easily by combining the ARCA/NTP mix, T7 RNA Polymerase Mix and a suitable DNA template. The kit also allows for partial incorporation of 5mCTP, Pseudo-UTP and other modified nucleotides into mRNA. After a brief DNase I treatment to remove the template DNA, capped mRNA is poly(A) tailed with Poly(A) Polymerase. 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 Kit (with tailing)Figure 2. Overview of mRNA synthesis work flow with the HiScribe T7 ARCA mRNA Kit (with tailing)

Kit Components

The following reagents are supplied with this product:

Store at (°C)

Concentration

ARCA/NTP Mix

-20

2X

DNase I (RNase-free)

-20

2 units/μl

Poly(A) Polymerase Reaction Buffer

-20

10X

LiCl Solution

-20

E. coli

Poly(A) Polymerase

-20

T7 RNA Polymerase Mix

-20

CLuc Control Template

-20

0.25 μg/μl


Properties and 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


Related Products

Companion Products


Notes
  1. All kit components should be stored at -20°C. The kit contains sufficient reagents for 20 reactions of 20 μl each. Each standard reaction yields up to 20 μg of capped mRNA from 1 μg control template. Up to 25 μg capped and tailed mRNA can be obtained after Poly(A) tailing and purification by LiCl precipitation.


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Safety Data Sheet

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








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 gbd@neb.com.

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.



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