Team:UNITN-Trento/Notebook/Labposts/08/37

From 2013.igem.org

{ "date" : "2013-08-14", "author" : "fabio-thomas", "title" : " blue light induction, NO WAY!!", "content" : " this morning I saw the results of the experiment and… horror: the induced bacteria pellet was lighter then the other two pellets (by the way, they weren’t blue, just a little bit bluish ) THAT SHOULDN’T HAVE HAPPENED!dang it.Maybe the blue LED was too intense, maybe there’s something wrong with our device, don’t know. We only need to take a crack at some other solution at the same time in order to solve this problem: first I will repeat the experiment with less intense blue light, then we are thinking about cotransforming bacteria with two plasmids, one with j23100-YF1-Fixj, the other with -FixK2-CI-Plambda-amilCPbecause we noticed that the device used yesterday lacks a terminator after fixJ and that could be the reason of the disfunction. Thomas made the cotransformation plates even though the plasmids actually aren’t compatible. Meanwhile we discussed about using a new backbone for the separate parts so that cotransformation could be achievable: so we made PCR amplification of the new vector pSB4K5, the first part (j23100-YF1-FixJ) and K592020 (FixK2-CI-Plambda-amilCP). As we can see from the gel, the screening of (j23100_YF1_FixJ) didn’t succeed, probably because the backbone’s structure is quite unusual.Furthermore I tried also to plate the entire device on 3 plates in order to make some mask experiment (induce colonies on the plate).", "tags" : " BlueLight_amilCP" }