Monday 29 August
Our last ligations haven't grown. Great. Let's start over...again. We don't know what we're doing wrong: the electrophoresis of the restrictions usually show the expected bands, and we've already tried 2 protocols for purification, and we've also made the final product more concentrated. However, the ligations or the transformation are still a barrier we always crash into. Deffinitely, synthetic biology is not standard, nor predictable.
At least we've found violet halos in our CV026 plates, what means two of the three AHLs we asked for are in good condition. Now, to test C12-3-oxo-HSL we have to seed another plate with C.violaceum with C6-3-oxo-HSL in order to generate a purple lawn in which we leave three drops of C12 at different concentrations. [To be continued]
Tuesday 30 August
Good news! Look at our fashionable plate! A perfect purple lawn with some white halos where the AHL was added, demonstrating it works perfectly.It's great when things work as expected for once...
However, the project is not going as well as we'd like, due to some reason we still don't understand. Anyway, time's running out, and let's face it, we obviously can't reach all the goals we planned in the beginning. A restructuration is indispensable.
Wednesday 31 August
Our new plan is to simplify the project as much as possible, this means giving up the idea of the full-adder and trying to build some basic logic gates instead. Though some of our planned ligations are still useful, we need some new DNA constructions, RNAi and stuff like that. So after hours of discussions about what logic gate to build and how, we start with the design of the sequences we want to ask for. Tedious work, but we need all our attention in order not to make any mistakes. This is crucial, all eyes are too few.
21.00 pm. Locked up in the lab because Carlos has gone home carrying the keys with him. Luckily, we have a projector here, so we don't panic: we watch another Game of Thrones episode while we wait for him to come back and rescue the rest of the team. This is really addictive!
Thursday 1 September
It almost makes us cry when we see how much the constructions will cost, but that's our final bet, so "Order placed" button and now we just can wait for a couple of weeks for C-Viral to send them. In the mean time we'll try tirelessly to get more ligations right so that we can do it properly with the constructiones once they're here.
No more Game of Thrones episodes to kill the time,we'll have to wait 'til April for the next season.
Friday 2 September
Not much to mention, the cycle has started again - for the fifth, tenth, hundredth time??. By this stage we can even repeat by heart the protocols for the plasmid extraction, calculate in 5 seconds the amounts of enzymes, buffer and water for the restriction mastermix and we could even prepare electrophoresis gels blindfolded and in record times.
In order to stop thinking about the amounts of money we've invested, Marta hosts a Roman party to celebrate Pedro and Ines' birthdays. Wine, grapes,laurel crowns, white togas, lyre chords, and as usual LOTS of exquisite food.