HiScribe® T7 ARCA mRNA Kit

Catalog #	Concentration	Size	List Price	Quantity	Your Price
E2065S		20 reactions	$513.00		$461.70

Catalog #	Size	List Price	Your Price
E2065S	20 reactions	$513.00	$461.70

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.

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 workflow with the HiScribe T7-ARCA mRNA Kit

Reagents Supplied

The following reagents are supplied with this product:

NEB #

Component Name

Component #

Stored at (°C)

Amount

Concentration

E2065S -20

T7 RNA Polymerase Mix	M0255AAVIAL	-20	1 x 0.04 ml
ARCA/NTP Mix	N2053AVIAL	-20	1 x 0.2 ml
DNase I (RNase-free)	M0303AAVIAL	-20	1 x 0.04 ml	2,000 units/ml
CLuc Control Template	N0247AVIAL	-20	1 x 20 µl	0.25 mg/ml
LiCl Solution	B2051AVIAL	-20	1 x 1.4 ml
Dithiothreitol (DTT)	B1222AVIAL	-20	1 x 0.5 ml	100 mM

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 (see Monarch^® RNA Cleanup Kits, NEB #T2040 or #T2050)
Gels, running buffers and gel box
Equipment for RNA analysis

Companion Products

Protocols

DataCards

manualE2065

FAQs

Citations

Quality Control Assay

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 Data Sheets

Legal And Disclaimer

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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.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.