Team:UNITN-Trento/Notebook/Labposts/08/14

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(Difference between revisions)

Latest revision as of 09:05, 3 October 2013

{ "date" : "2013-08-06", "author" : "bruno-viola-caterina-gabriele-thomas", "title" : "Surrender now or prepare to fight, Meowth! That's right.", "content" : "Today we made 5 different competent cells following the transformation protocol:
- NEB10β
- TB1
- TOP10
- JM109
- BL21

Then we trasformed in these cells R0010-K592009 in psB1C3 and K1065300.

", "tags" : "cells competent" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-08-07",
+	"date" : "2013-08-06",
-	"author" : "thomas",
+	"author" : "bruno-viola-caterina-gabriele-thomas",
-	"title" : "pXil + GFP transformation in B. subtilis",
+	"title" : "Surrender now or prepare to fight, Meowth! That's right.",
-	"content" : "<html>In order to transform <i>Bacillus</i> with pXyl (BBa_K823024) + GFP (BBa_E0840), reamplified and screened <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-08-02-thomas\">the 2nd of August</a>, I prepared an inoculum in 2 ml of MN completed medium (<a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#subtilis-transformation\">see protocol</a>). While I was waiting that O.D.600 reached 0.5, I proceeded digesting 2,5&micro;g of sample 2 and sample 3 of the construct pXyl + GFP using ScaI as restriction enzyme. I did this in order to linearize the plasmid and allow his integration in <i>B. subtilis</i> genome. 0.5&micro;g of each sample were then run on an agarose gel for screening.</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/9/90/Tn-2013_gel_pXyl%2BGFP_ScaI_digested.JPG\" width=\"450px\" /></center></html>}}<html> As you can see from the picture, there are two not expected bands: one at height of 4500 bp and one at height of about 1800 bp. I can't explain this two bands since ScaI should digest only at only one sites on the construct.. However there's also a band at the height between 5000 and 6000 bp as expected (pXyl+GFP = 5782 bp) so I transformed it into <i>bacillus</i> anyway.</html>",
+	"content" : "<html>Today we made 5 different competent cells following the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Competent-cells-transformation\">transformation protocol</a>:<br/>- NEB10&beta;<br/>- TB1<br/>- TOP10<br/>- JM109<br/>- BL21<br/><br/>Then we trasformed in these cells R0010-K592009 in psB1C3 and K1065300. <br/><br/><br/><br/><br/><br/><br/><br/></html>",
-	"tags" : "pXyl-GFP-B. subtilis"
+	"tags" : "cells competent"
 }