Codon optimized firefly luciferase was cloned into an mRNA production plasmid (optimized 3′ and 5′ UTR and containing a 101 polyA tail), in vitro transcribed in the presence in the presence of N1-methylpseudouridine modified nucleoside (N1mψ), co-transcriptionally capped using the CleanCap™ technology (TriLink) and cellulose purified to remove dsRNA. Purified mRNA was ethanol precipitated, washed, resuspended in nuclease-free water, and subjected to quality control (electrophoresis, dot blot, and transfection into human dendritic cells). mRNA was stored at −80°C until use32 (link).
Protocol detail
Firefly Luciferase mRNA Production
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
5 utr
Cellulose
Codon
Dendritic cells
Dot blot
Dsrna
Electrophoresis
Ethanol
Firefly luciferase
Human
Mrna
N1 methylpseudouridine
Nucleoside
Plasmid
Polya tail
Transfection
Corresponding Organization : California Institute for Regenerative Medicine
Other organizations : Gdańsk Medical University
Top 5 similar protocols
Variable analysis
independent variables
- Codon optimized firefly luciferase
- N1-methylpseudouridine modified nucleoside (N1mψ)
- CleanCap™ technology (TriLink)
- Quality control (electrophoresis, dot blot, and transfection into human dendritic cells)
- MRNA production plasmid (optimized 3′ and 5′ UTR and containing a 101 polyA tail)
- In vitro transcription
- Cellulose purification to remove dsRNA
- Ethanol precipitation, washing, and resuspension in nuclease-free water
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!