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mMESSAGE mMACHINE™ T7 mRNA Kit with CleanCap™ Reagent AG, 1000 rxns - FAQs

mMESSAGE mMACHINE™ T7 mRNA Kit with CleanCap™ Reagent AG, 1000 rxns - FAQs

View additional product information for mMESSAGE mMACHINE™ T7 mRNA Kit with CleanCap™ Reagent AG - FAQs (A57621, A57620-25, A57620)

8 product FAQs found

How is CleanCap Reagent AG different from ARCA (Anti-Reverse Cap Analog) technology?

CleanCap is a new capping technology that can provide higher mRNA yields and capping efficiencies than ARCA. CleanCap contains the cap 1 structure, which is found in higher eukaryotes such as mice or humans. ARCA contains the cap 0 structure that is found in lower eukaryotes such as yeast. Cap 1 mRNA can have superior translational activities in humans compared to Cap 0 mRNAs.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

How can I introduce the RNA synthesized using the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621), into cells?

The RNA can be introduced into cells by electroporation or transfection. Our Neon NxT Electroporation System can be used to introduce the RNA by electroporation. Our Lipofectamine MessengerMAX Transfection Reagent is designed for mRNA delivery and works with many cell types.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

How can I purify the RNA synthesized using the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621)?

The RNA synthesized using the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621) can be purified using the provided LiCl Precipitation Solution. We recommend referring to the user guide for details. Alternatively, the RNA can be purified using spin columns such with the MEGAclear Transcription Clean-Up Kit (Cat. No. AM1908). Note that the MEGAclear Transcription Clean-Up Kit can purify up to 500 µg RNA and that it only purifies RNAs >100 nucleotides. If shorter RNAs need to be purified, we recommend using the GeneJET RNA Cleanup and Concentration Micro Kit (Cat. No. K0841).

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Can I use modified nucleotides with the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621)?

Yes, modified nucleotides can be used. We recommend referring to the user guide for guidance on this.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

With the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621), do you recommend using a template encoded poly-A tail?

Yes, we recommend using a template encoded poly-A tail as it yields better expression than when a poly-A tail is added enzymatically using poly-A polymerase. Refer to the user guide for guidance on making a template with a poly-A tail.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

With the mMESSAGE mMACHINE T7 mRNA Kit with CleanCap Reagent AG (Cat. No. A57620, A57621), do you recommend using a linearized plasmid or a PCR product as the template?

Both types of templates are suitable for making mRNA with these kits and it is a matter of preference as to which one is used. We find that it is easier to get the amounts of template needed for large scale reactions (e.g., 1 mg mRNA or more) when using linearized plasmid as the template.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.

Do I need to change my promoter sequence to use CleanCap Reagent AG?

Yes, the promoter sequence should be modified. CleanCap Reagent AG needs an AG initiating promoter to achieve high capping efficiencies (>90%). Refer to the user guide for guidance on how to design the promoter so that it is compatible with CleanCap Reagent AG.

Find additional tips, troubleshooting help, and resources within our Nucleic Acid Purification and Analysis Support Center.