Team:Evry/Notebook/Transposons/13.10.14

From 2014.igem.org

(Difference between revisions)
(Created page with "<html> <div class="cd-timeline-block"> </html> {{:Team:Evry/Notebook/CellCharacterization/Transformation/TopImage}} <html> <div class="cd-timeline-content"> <p align="just...")
 
(10 intermediate revisions not shown)
Line 6: Line 6:
     <div class="cd-timeline-content">
     <div class="cd-timeline-content">
-
<p align="justify">
 
-
== BBa_1413044 retrieval of the transposon plasmid==
 
 +
<h4> BBa_1413044 retrieval of the transposon plasmid </h4>
 +
<br><p>
 +
strategy:</p>
 +
<div align="center">
 +
<br><img src="https://static.igem.org/mediawiki/2014/3/33/Evry_Strat1.jpg" width="75%"/></div>
 +
<br>
 +
<p>
Digestion of BBa_1413044 by BglII :
Digestion of BBa_1413044 by BglII :
-
50 ul final volume, 5 ul Buffer NeB 3.1, 5 ul (20 ng/ul) BBa_1413044 in pSB1C3, 1 ul BglII, 39 ul H2O.
+
<br>50 ul final volume, 5 ul Buffer NeB 3.1, 5 ul (20 ng/ul) BBa_1413044 in pSB1C3, 1 ul BglII, 39 ul H2O.
-
 
+
<br>
Gel electrophoresis:
Gel electrophoresis:
-
 
+
<br><div align="center">
-
GEEEEEEEEEEEEEEEEEEEEEELLLLLLLLLL
+
<img src="https://static.igem.org/mediawiki/2014/3/31/Evry_Bba_transpo.JPG" width="25%"/></div>
-
<img src="" width="50%"/>
+
<br>
-
 
+
Purification sur gel:
Purification sur gel:
-
 
+
<br>
Kit thermoscientific "Gel extraction". Elution final in 30 ul.
Kit thermoscientific "Gel extraction". Elution final in 30 ul.
-
 
+
<br></p>
-
== Obtaining the Kan gene on pNK2 ==
+
<h4>Obtaining the Kan gene on pNK2</h4>
-
 
+
<br><p>
-
PCR: enzyme: Q5; template: 150 ng pNK2; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s
+
PCR: enzyme: Q5; template: 150 ng pNK2 first three well from new stock, next ones the old stock; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s
-
 
+
<br></p><div align="center">
-
<img src="" width="50%"/>
+
<img src="https://static.igem.org/mediawiki/2014/9/90/Evry_Kan_extract.JPG" width="40%"/></div>
-
 
+
<br><p>
Purification sur gel:
Purification sur gel:
-
 
+
<br>
Kit thermoscientific "Gel extraction". Elution final in 30 ul.
Kit thermoscientific "Gel extraction". Elution final in 30 ul.
-
 
+
<br></p>
-
 
+
-
<br>.
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+
-
 
+

Latest revision as of 22:41, 17 October 2014

Picture

BBa_1413044 retrieval of the transposon plasmid


strategy:



Digestion of BBa_1413044 by BglII :
50 ul final volume, 5 ul Buffer NeB 3.1, 5 ul (20 ng/ul) BBa_1413044 in pSB1C3, 1 ul BglII, 39 ul H2O.
Gel electrophoresis:


Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.

Obtaining the Kan gene on pNK2


PCR: enzyme: Q5; template: 150 ng pNK2 first three well from new stock, next ones the old stock; oligo: F 82/ R 83; Tm tested: 55/60/65; elongation time: 1m30s


Purification sur gel:
Kit thermoscientific "Gel extraction". Elution final in 30 ul.

Oct 13

Retrieved from "http://2014.igem.org/Team:Evry/Notebook/Transposons/13.10.14"