How to Perform an in vitro transcription experiment

The Department of Biological Sciences at the University of Alberta brings you this flash animated video tutorial specifically relevant to your student laboratory courses, specifically microbiology. If you're a student at any school of biology, this information will be helpful for learning how to perform in vitro transcription.

In this science experiment, the protein products of a specific plasmid (or other suitable genetic material) are isolated, analyzed, and identified. The experiment is "in vitro" since a modified cell lysate, the S30 extract, is used to carry out transcription and translation.

This extract has been treated to degrade DNA and mRNA from the lysed cells it came from. To this, a circular plasmid is added, along with amino acids. For detection methods, one amino acid is radiolabeled. The reaction can be sampled to determine reaction rate analysis, and the remaining reaction mixture is characterized for protein, usually by SDS-PAGE.