Team:UNITN-Trento/Notebook/Labposts/07/58

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(Difference between revisions)

Latest revision as of 10:46, 3 October 2013

{"date" : "2013-07-24","author" : "thomas","title" : "Screening of AraC-pBAD + EFE + Venus","content" : "This is the part five of this experiment. I proceeded with a miniprep purification of the previously prepared inocula and a screening digestion in order to verify the construct.

Quantification Results

Sample	Concentration
A from ligation 1:1	852.6 ng/ul
B from ligation 1:1	1123 ng/ul
C from ligation 1:4	685.8 ng/ul
D from ligation 1:4	2128.3 ng/ul
E from ligation 1:1	1760 ng/ul
A from ligation 1:1	852.6 ng/ul

Gel Image

As you can see from the gel image, only one sample seems to confirm our fusion protein and will be sent for sequencing. Lane 4: insert 3024 bp, vector 2070 bp. Kapa universal ladder was adopted.","tags" : "EFE-Venus"}

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+{"date" : "2013-07-24","author" : "thomas","title" : "Screening of AraC-pBAD + EFE + Venus","content" : "<html>This is the part five of <a href=\"https://2013.igem.org/wiki/index.php?title=Team:UNITN-Trento/Notebook#tn-post-2013-07-18-thomas-michele\">this experiment</a>. I proceeded with a <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\">miniprep purification</a> of the previously prepared inocula and a <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">screening digestion</a> in order to verify the construct. <br/><br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Quantification Results|<html><center><TABLE CLASS=\"tn-sp-table\"><tr><th>Sample</th><th>Concentration</th></tr><tr><td>A from ligation 1:1 </td><td>852.6 ng/ul</td></tr><tr><td>B from ligation 1:1 </td><td>1123 ng/ul</td></tr><tr><td>C from ligation 1:4 </td><td>685.8 ng/ul</td></tr><tr><td>D from ligation 1:4</td><td>2128.3 ng/ul</td></tr><tr><td>E from ligation 1:1 </td><td>1760 ng/ul</td></tr><tr><td>A from ligation 1:1 </td><td>852.6 ng/ul</td></tr></TABLE></center></html>}}<html> <br/></html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|Gel Image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/d/d9/Tn-2013_venus_screening.JPG\" style =\"width: 450px\"></center></html>}}<html> As you can see from  the gel image, only one sample seems to confirm our fusion protein and will be sent for sequencing. Lane 4: insert 3024 bp, vector 2070 bp. Kapa universal ladder was adopted.</html>","tags" : "EFE-Venus"}
-	"date" : "2013-07-28",
-	"author" : "thomas",
-	"title" : "EFE + Venus Expression test: FAILED!",
-	"content" : "<html>Starting from an inoculum, I diluted the cells 1:100 in fresh LB and I waited until O.D.600 of the sample reached 0.5. I induced then the cells (V=5ml) with 25 ul of Arabinose (5mM) and I incubated them into thermoshaker at 37&deg;C for 4 hours. One sample were not induced and used as negative control.After 4 hours I tried to use the trans-UV too see any differences between induced sample and control. Both culture had the same color (not fluorescence) so the experiment failed. However the sample was sent for sequencing so now I'll wait the result before restart all the cloning steps..</html>",
-	"tags" : "EFE-Venus"
-}