Takara IVTproTM T7 mRNA Synthesis Kit

Description

Takara IVTpro mRNA Synthesis Kit enables the production of large amounts of mRNA through in vitro transcription using an optimized, highly efficient T7 RNA polymerase. The kit includes separate NTPs for easy optimization or replacement with modified nucleosides, such as pseudouridine. The kit synthesizes up to 200 mug or more of mRNA per 20 mul reaction, includes DNase I to digest the template DNA after the in vitro transcription reaction and a lithium chloride LiCl solution to purify the synthesized mRNA for downstream applications. The kit also includes a Positive Control Template FLuc plasmid containing the T7 promoter, transcription initiation sequence AGG , 5- and 3-UTRs, FLuc CDS, and poly A sequence. Easily scale up to 10-fold 200 mul reaction volume without affecting the expected mRNA yield. Our products are to be used forResearch Use Only. They may not be used for any other purpose, including, but not limited to, use in humans, therapeutic or diagnostic use, or commercial use of any kind. Our products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without our prior written approval. Back Addition of the CleanCap Reagent AG 3 OMe in IVT reaction has no significant effect on mRNA yield Panel A . HEK293T cells transfected with mRNA containing a cap structure show significatly higher protein expression compared to cells transfected with mRNA without a cap Panel B . Back The mRNA yield was proportional to the reaction volume up to 200 mul with no significant change in mRNA concentration Panel A . Scaling up of the IVT reaction volume does not affect the quality of RNA product Panel B . Back Purification of RNA product using LiCl was comparable to purification with NucleoSpin column when two elutions were performed Panel A . HEK293T cells transfected with mRNA purified using LiCl shows protein expression comparable to mRNA purified using NucleoSpin columns Panel B . Back The IVT reaction was performed using various sizes of DNA Templates with CleanCap Reagent AG 3 OMe and N1-methyl pseudo UTP. Each resulting mRNA 200 ng was denatured at 65 C for 10 min, followed by analysis using 1.2 % formaldehyde gel electrophoresis. Back