HiScribe™ T7 ARCA mRNA Kit
- Generate up to 20 μg of capped mRNA per reaction
- mRNA capping, DNA removal, mRNA tailing and purification complete in 2 hrs. 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 with tailing reagents
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. 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.
The following reagents are supplied with this product:
Store at (°C) Concentration LiCl Solution -20 DNase I (RNase-free) -20 2 units/μl T7 RNA Polymerase Mix -20 CLuc Control Template -20 0.25 μg/μl ARCA/NTP Mix 2 X
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