K1063-25 | HyperScribe™ All in One mRNA Synthesis Kit (ARCA, T7, poly(A))

Product Citation

1. Ning Luan, Han Cao, et al. "Comparison of Immune Responses between Inactivated and mRNA SARS-CoV-2 Vaccines Used for a Booster Dose in Mice." Viruses. 2023 Jun 11;15(6):1351. PMID: 37376650

Description

HyperScribe™ All in One mRNA Synthesis Kit (ARCA, T7, poly(A)) is designed for quick production of ARCA capped and poly(A) tailed mRNA in vitro. Capped mRNA is synthesized by co-transcriptional incorporation of Anti-Reverse Cap Analog (ARCA, Catalog No. B8175) using T7 RNA Polymerase. The capping of ARCA ensures high translation efficacy. After a brief DNase I treatment to remove the template DNA, capped mRNA is poly(A) tailed with Poly(A) Polymerase. Polyadenylation (i.e. addition of Poly(A) tail) plays an important role in the stabilization of RNA in eukaryotes and enhances the efficiency of translation initiation.

RNA synthesized using this kit has many applications in biological experiments, such as in vitro translation, antisense RNA and RNAi experiments, RNA vaccines, RNA structure and function studies, ribozyme biochemistry, RNase protein experiments and probe-based hybridization blots.

The kit contains sufficient reagents to carry out 25 reactions, 20 μL each time. Up to 50 μg of RNA can be generated with 1 μg of control template per standard reaction.

An upgraded version of this product is available with higher yield (~100 μg), please check out K1406 HyperScribe™ Co-transcription mRNA Synthesis Kit Plus (ARCA, T7) for more details. (Note: it does not include the reagents for the poly A tailing, please incorporate the poly A sequence in your plasmid or primer).

Quality Control

Protocol

Components and Storage

(1) Capped RNA Synthesis with ARCA
Components	25 rxns
T7 RNA Polymerase Mix	50 μL
10X Reaction Buffer	50 μL
ATP (20 mM)	50 μL
UTP (20 mM)	50 μL
CTP (20 mM)	50 μL
GTP (20 mM)	37.5 μL
ARCA (60 mM)	50 μL
Control Template (0.5 μg/μL)	5 μL
RNase-free H₂O	0.5 mL
Store all kit components at -20°C.
(2) Tailing Reaction
Components	25 rxns
E-PAP (2 units/μL)	100 μL
5X E-PAP Buffer	600 μL
ATP Solution (100 mM)	100 μL
25 mM MnCl₂	250 μL
Nuclease - free Water	1 mL x 2
Store all kit components at -20°C.

Storage

Store all kit components at -20°C.

HyperScribe™ All in One mRNA Synthesis Kit (ARCA, T7, poly(A)) [Used for the synthesis of ARCA capped and poly(A) tailed mRNA in vitro]

Cat# K1063-25

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Brand : APExBIO Technology