HyperScribe™ All in One mRNA Synthesis Kit Plus 1 (ARCA, 5mCTP, ψUTP, T7, poly(A))
HyperScribe™ All in One mRNA Synthesis Kit Plus 1 (ARCA, 5mCTP, ψUTP, T7, poly(A)) is designed for quick production of ARCA capped, 5mCTP and ψUTP modified, 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. The addition of 5mCTP (Catalog No. B7967) / ψUTP (Catalog No. B7972) can reduce host cell immune response. After a brief DNase I treatment to remove the template DNA, capped and modified 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 K1407 HyperScribe™ Co-transcription mRNA Synthesis Kit Plus (ARCA, 5mCTP, ψUTP, 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).(1) RNA Synthesis with ARCA, 5mCTP and ψUTP (Pseudo-UTP) | |
| Components | 25 rxns |
| T7 RNA Polymerase Mix | 50 μL |
| 10X Reaction Buffer | 50 μL |
| ATP (20 mM) | 50 μL |
| UTP (20 mM) | 25 μL |
| CTP (20 mM) | 25 μL |
| GTP (20 mM) | 25 μL |
| ARCA (60 mM) | 35 μL |
| 5mCTP (10 mM) | 50 μL |
| Pseudo-UTP (10 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. | |
