Team:UNITN-Trento/Notebook/Labposts/05/1

From 2013.igem.org

(Difference between revisions)
(Undo revision 78353 by Ggirelli (talk))
Line 1: Line 1:
{
{
-
"date" : "2013-05-16",
+
"date" : "2013-08-14",
-
"author" : "caterina-gabriele"
+
"author" : "fabio-thomas",
-
"title" : "First day at CIBIO",
+
"title" : "blue light induction, NO WAY!!",
-
"content" : "First day as iGEMMers at CIBIO (<i>Center for the Integrative Biology</i>)!<br/>
+
"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).",
-
We extracted <b>BBa_J45319</b>, <b>BBa_J45320</b>, <b>BBa_J45004</b> and <b>BBa_J45119</b> and then transformed them into NEB10&beta; cells.",
+
"tags" : "j23100_YF1_FixJ_FixK2_CI_Plambda_amilCP"
-
"tags" : "PchA-PchB-PchBA-BSMT1"
+
}
}

Revision as of 12:20, 19 August 2013

{ "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" : "j23100_YF1_FixJ_FixK2_CI_Plambda_amilCP" }