Team:UNITN-Trento/Notebook/Labposts/08/26

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(Difference between revisions)

Latest revision as of 09:10, 3 October 2013

{ "date" : "2013-08-11", "author" : "thomas-viola", "title" : "Overnight cultures", "content" : "Starting with the plates we did yesterday, we prepared the following overnight cultures.

Overnight cultures prepared

Plate	N° of colonies taken
S04617 + EFE ligation 1:2	3
AraCpBAD - EFE ligation 1:1	2
AraCpBAD - EFE ligation 1:2	2

", "tags" : "pFixK-cI-pLambda-EFE-AraCpBAD" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-08-12",
+	"date" : "2013-08-11",
 	"author" : "thomas-viola",
-	"title" : "We don't give up!",
+	"title" : "Overnight cultures",
-	"content" : "<html>Ok yesterday we discovered that the ligation between <a href=\"http://parts.igem.org/Part:BBa_S04617\">BBa_S04617</a> and <a href=\"http://parts.igem.org/Part:BBa_K1065002\">BBa_K1065002</a> failed so today we focused on a new cloning strategy. We decided to amplify both parts by PCR following <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#OneTaq-Phu-PCR\">this protocol</a>. BBa_Fwd and BBa_Rev primers were adopted. PCR products were then screened, purified and quantified at the nanodrop.. </html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/3/36/Tn-2013_gel_PCR_s04617_K1065002.jpg\"></center></html>}}<html> As you can see from the picture, the gel confirmed our PCR reaction since in lane 1 there is a band at heigh of 1215 bp (BBa_K1065002) and in lane 3 one band at heigh of 1245 bp (BBa_S04617). Kapa universal ladder was adopted.We proceeded then performing an o/n digestion on the following parts in order to do a tri Assembly:<center><table><tr><th>Part</th><th>Enzymes used</th><th>quantification after purification</th></tr><tr><td>pSB1C3</td><td>EcorI+PstI</td><td>90.1 ng/&micro;l</td></tr><tr><td>EFE+B0015</td><td>XbaI+PstI</td><td>104.7 ng/&micro;l</td></tr><tr><td>s04617</td><td>EcorI+SpeI</td><td>133.5 ng/&micro;l</td></tr></table></center>Tomorrow we will proceed with a ligation of the three digestion products..!</html>",
+	"content" : "<html>Starting with the plates we did <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-08-10-viola-thomas\">yesterday</a>, we prepared the following overnight cultures. </html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Overnight cultures prepared|<html><center><table><tr><th>Plate</th><th>N° of colonies taken</th></tr><tr><td>S04617 + EFE ligation 1:2</td><td>3</td></tr><tr><td>AraCpBAD - EFE ligation 1:1 </td><td>2</td></tr><tr><td>AraCpBAD - EFE ligation 1:2</td><td>2</td></tr></table></center></html>}}<html> </html>",
-	"tags" : "pFixK-cI-pLambda-EFE"
+	"tags" : "pFixK-cI-pLambda-EFE-AraCpBAD"
 }