This week we will be going over the following subjects:
IVT Process Overview:
- Components: DNA template, RNA polymerase, nucleotides, and buffers.
- Reaction conditions: Temperature, pH, and ionic strength.
- Optimization strategies for yield and quality.
mRNA Capping and Polyadenylation:
- Co-transcriptional capping methods.
- Poly(A) tail addition techniques.
- Impact on mRNA stability and translation efficiency.
Purification of mRNA:
- Removal of DNA template and enzymes.
- RNA cleanup methods: Column-based and precipitation techniques.
- Quality control: Integrity assessment via gel electrophoresis and spectrophotometry.
Laboratory Work Real-World Application:
- Example: IVT production of mRNA vaccines for COVID-19.