HiScribe™ T7 ARCA mRNA Kit (with tailing)
- Generate up to 25 μg of capped and tailed mRNA per reaction
- mRNA capping, DNA removal, mRNA tailing and purification complete in 2 hours. 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 without tailing reagents (NEB #E2065)
Avoiding RNase Contamination
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.
The following reagents are supplied with this product:
Store at (°C) Concentration Poly(A) Polymerase Reaction Buffer -20 10 X LiCl Solution -20 T7 RNA Polymerase Mix -20 DNase I (RNase-free) -20 2 units/µl E. coli Poly(A) Polymerase -20 CLuc Control Template -20 0.25 µg/µl ARCA/NTP Mix -20 2 X
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