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Team:Cambridge/Diary/Week 14
From 2012.igem.org
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Contents |
Monday
Andreas fully tested the instrumentation today. A dilution series was made to test the sensitivity of the sensor in the right geometry. More details can be found in the lab-book.
Still no successful ligations for the biobricks, as a couple of colony PCRs show. Looks like the insert for the SpoVA biobrick may not have been digested. Oops...
The platereader also gets a thorough workout, as we try one last assay on our magnesium riboswitch construct. 24 hours later, we might get some nice data.
We persevere though. While some of us concentrate on making the wiki presentable, Emmy and Oli (though mostly Emmy) set up 5 more biobricking restriction ligation reactions. Bands from the gels are beautifully pure, so we're hoping for at least one success.
Alas, the transformants only get plated out at 11 am.
Tuesday
Days have become a bit arbitrary now, haven't they? The last entry only finished at 11 o'clock.
Andreas tested the instrumentation for colour recognition. More details can be found in the Lab book. Wiki dominates proceedings from now on, unfortunately (or fortunately? It's certainly easier to do than ligations). Emmy seems keen to sacrafice her own well being for the success of the wiki, having stayed up all night working on t-shirt designs and then staying until midnight to get some more design work done. She finally goes home after a 36 hour shift - I doubt any of the rest of us can really have reason to complain.
Unfortunately, we still haven't got any colonies on the ligation plates. On the bright side, there aren't any on the positive transformation control plate either, so perhaps not so desperate as we feared. We'll have to be patient and see what happens tomorrow.
And Jolyon sets out to try the Miller assay one last time. Even though it worked pretty well last time, we'd still like some more data to average out.
Wednesday
The final day is here, and we're dedicated to getting the wiki as presentable as possible. Thank God Emmy hasn't been stricken by some disease that strikes down the poorly rested.
Not that wet work has completely passed us by, on this, our final day. As I write, a plate reader assay is running to check if Ratiometrica works as expected. On top of this, Stewart is taking a time lapse series of photos to see if our promoter speeds up germination as expected. See our final characterization data on the results page, as well as the respective pages for each of our parts.
Plus, we finally got some colonies on the ligation plates! Unfortunately, colony PCR with the sequencing primers reveals that all we have is recircularized backbone. We don't even know how it managed to do this, because we treated the vector with alkaline phosphatase. It seems that the ligation fairy is simply not smiling on us.
Thursday
Well. I did say yesterday was our final day. But you know, Wiki-ing is just so addictive, it's impossible to shut it out until the last possible moment. So this is it - the final hours. It's been a ridiculous ride over the last three months, and even though some of our experiments didn't go exactly to plan (I'm looking at you, platereader, destroyer of dreams) it's impossible to deny that we've learnt a great deal. So, nothing left to say but - See you in Amsterdam!
