Team:UNITN-Trento/Notebook/Labposts/07/53

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Revision as of 08:39, 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" }

@@ Line 1: / Line 1: @@
 {
-	"date" : "2013-07-10",
+	"date" : "2013-07-24",
-	"author" : "emil",
+	"author" : "thomas",
-	"title" : " Purification of Inocula(09/7): the final countdown",
+	"title" : "Screening of AraC-pBAD + EFE + Venus",
-	"content" : "<html>I (yes I'm egocentric) did the miniprep(wath a new thing!) of the inocula(I didn't purified the 1:2 due to their despicable red color) previously done(the last I hope) following(is always the same but I have to write it anyway)the <a href=\"https://2013.igem.org/Team:UNITN-Trento/Protocols#miniprep\">miniprep protocol</a>(OOOH!that's incredible!).These are(obviously)the results of the quantification:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|results|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Quantities</th></tr><tr><td>BBa_K823026+BBa_E0840(1:1) </td><td> 421.3ng/&micro;l</td></tr><tr><td>BBa_K823026+BBa_E0840(1:1) </td><td> 438.6ng/&micro;l</td></tr>tr><td>BBa_K823026+BBa_E0840(1:3) </td><td> 356.7ng/&micro;l</td></tr><tr><td>BBa_K823026+BBa_E0840(1:3) </td><td> 415.2ng/&micro;l</td></tr><tr><td>BBa_K823024+BBa_E0840 </td><td> 401.2ng/&micro;l</td></tr>tr><td>BBa_K823024+BBa_E0840 </td><td> 365ng/&micro;l</td></tr></table></center></html>}}<html>Afterwards I screened them(800 ng for each sample) following the <a href =\"https://2013.igem.org/Team:UNITN-Trento/Protocols#Digestion\">digestion protocol</a> so I have obtained these results:</html>{{:Team:UNITN-Trento/Templates/Styles/Spoiler|gel order|<html><center><table class=\"tn-sp-table\"><tr><th>Sample</th><th>Well</th></tr>                <tr><td>ladder </td><td>1</td></tr><tr><td>BBa_K823026+BBa_E0840(1:1) </td><td>2</td></tr><tr><td>BBa_K823026+BBa_E0840(1:1) </td><td>3</td></tr>tr><td>BBa_K823026+BBa_E0840(1:3) </td><td>4</td></tr><tr><td>BBa_K823026+BBa_E0840(1:3) </td><td>5</td></tr><tr><td>BBa_K823024+BBa_E0840 </td><td>6</td></tr>tr><td>BBa_K823024+BBa_E0840 </td><td>7</td></tr></table></center></html>}}{{:Team:UNITN-Trento/Templates/Styles/Spoiler|image|<html><center><img src=\"https://static.igem.org/mediawiki/2013/1/15/Tn-2013_DSC0161.JPG\" width=\"450px\"></html>}}<html>As we can see  in the last 4 samples the lower band is clearly at 1000 bp then the GFP is probably present, the  succesfull samples were sent to be sequenced!</html>",
+	"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" : "pSpac-PXyl-GFP"
+	"tags" : "EFE-Venus"
 }