In this lab we made a reaction in an RNAse free environment that synthesised GFP and FIAsH proteins.
synthetic cell: "An artificial cell or minimal cell is an engineered particle that mimics one or many functions of a biological cell. The term does not refer to a specific physical entity, but rather to the idea that certain functions or structures of biological cells can be replaced or supplemented with a synthetic entity."
Master mix: a premixed concentrated solution that has all of the components for a reaction that are not sample-specific.
The first part of this lab was figuring out the concentration and amounts of everything that needed to go into the synthetic cell. This was a lot of massaging the numbers in an excel spread sheet, followed by a lot of pipetting. As we needed the final volume to be 20ul, but there were 10+ compenents to each mixture, we ran into the limitations of our pipettes, which can only measure down to 2uL and had to do several pre-mixing steps to acheive the correct final concentrations.
Each person in the group made one mix for producing GFP and one for FIAsH proteins, though we took these samples from the master mix, so they really should all be the same. We put them in the clear, flat bottom well plate and measured their fluorescence after 12 hours of incubation at 37C. The settings were: Excitation: 470nm, Emission: 515nm, Gain: 3000. We measured 2.3-2.6x higher fluorescence in the GFP and FIAsH samples respectively in comparison to measurements taken on empty wells.