Takara IVTpro mRNA Synthesis System

Takara IVTpro mRNA Synthesis System includes all components necessary for DNA template preparation and high-yield in vitro transcription reaction. This kit includes:

Cloning Kit for mRNA Template (Cat. # 6143)

Cloning Kit for mRNA Template enables easy cloning of a template DNA into a pre-linearized plasmid (included in the kit) for downstream in vitro transcription reactions. The pre-linearized plasmid contains the T7 promoter, transcription initiation sequence (AGG), 5'- and 3'-UTR (untranslated region), and poly(A) sequence. The kit also includes an In-Fusion Snap Assembly Master Mix for seamless cloning of the target DNA into the pre-linearized vector and a FLuc Control Fragment (consisting of a 15 bp, 3' In-Fusion sequence; Fluc CDS; and 15 bp, 5' In-Fusion sequence).

Takara IVTpro mRNA Synthesis Kit (Cat. # 6144)

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 μg or more of mRNA per 20 μl 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 FLuc positive control template 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 μl reaction volume) without affecting the expected mRNA yield.

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Affect of CleanCap Reagent AG (3’ OMe) on RNA yield and the expression of FLuc mRNA

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).

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Increasing IVT reaction volume has no effect on RNA yield or quality.

The mRNA yield was proportional to the reaction volume up to 200 µl 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).

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Effect of different RNA purification methods on RNA yield and protein expression.

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).

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6141: Takara IVTpro mRNA Synthesis System

Cloning Kit for mRNA Template enables easy cloning of a template DNA into a pre-linearized plasmid (included in the kit) for downstream in vitro transcription reactions. The pre-linearized plasmid contains the T7 promoter, transcription initiation sequence (AGG), 5'- and 3'-UTRs (untranslated region), and poly(A) sequence. The kit also includes an In-Fusion Snap Assembly Master Mix for seamless cloning of the target DNA into the pre-linearized vector and a FLuc Control Fragment, consisting of 15 bp 3' and 5' In-Fusion sequences and Fluc CDS (1,683 bp).

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6143: Cloning Kit for mRNA Template

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 μg or more of mRNA per 20 μl 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 μl reaction volume) without affecting the expected mRNA yield.

Back

Affect of CleanCap Reagent AG (3’ OMe) on RNA yield and the expression of FLuc mRNA

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

Increasing IVT reaction volume has no effect on RNA yield or quality.

The mRNA yield was proportional to the reaction volume up to 200 µl 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

Effect of different RNA purification methods on RNA yield and protein expression.

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).

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6144: Takara IVTpro T7 mRNA Synthesis Kit