01 / The question
How can you tell that you made the right protein?
The project followed production of fluorescent KillerRed in E. coli through to characterisation. Producing coloured material and demonstrating protein identity are different tasks.
02 / The intuition
A mechanism worth testing.
Use several readouts with different failure modes. A purification fraction should be assessed by its composition and identity, not only its colour.
03 / The work
Inside the method.
Explore each part of the approach.
01Build and express
The documented work covered cloning checks, plasmid preparation and recombinant expression.
02Separate the protein
Affinity purification and fraction collection were used to enrich the target protein.
03Cross-check identity
SDS-PAGE, Western blot, fluorescence and quantitative assays provided complementary characterisation.
04 / The observations
What emerged.
The notes describe protein recovery and multiple identity checks. They also report a low target-protein fraction, making purity an important part of the story rather than a detail to omit.
This account is based on recorded project notes. Original reports, figures and datasets are not embedded here.
05 / The limits
Where the evidence stops.
Yield and purity require the original gels, calibration curves and fraction records. The present account is a process summary, not a validated production specification.
The academic foundations