Team:Warwick/Microscopy
Fluorescence microscopy
In order to visualise our cells and determine whether the modified zinc finger protein was binding to the surface of our E. coli cell, we also included a FLAG tag domain in the protein.
This allowed us to use anti-FLAG antibodies to conduct an immuno-fluorescence test to see the location of the protein in the cells.
Reciprocal Test
After determining that our modified zinc fingers were being expressed on the surface of the E. coli cell, we set out to prove whether our modified cell could bind DNA discriminately. If the cell binds DNA, but any old piece of DNA binds instead of the DNA we have selected, this would go against the aims of our project.
We set out to prove this using fluorescein bound oligonucleotides that contained our zinc finger target DNA sequence. By fixing different cells to slides and washing them with different oligos we could determine whether the zinc fingers bind only target DNA, or any DNA.