Team:Bordeaux/Fourth

From 2012.igem.org

(Difference between revisions)
(Created page with "<html lang="en"> <head> <title>Note Book - iGEM Bordeaux 2012</title> <meta charset="utf-8"> <link rel="stylesheet" href="http://cdjemiel.free.fr/igem2012/css/reset.css" type=...")
Line 62: Line 62:
</p>
</p>
                                               <div class="img-box">
                                               <div class="img-box">
-
<figure><a href="#"><img src="http://openwetware.org/images/a/a7/NewBB2307.jpg" alt=""></a></figure>
+
<figure><a href="#"><img src="https://static.igem.org/mediawiki/2012/a/a7/NewBB2307.jpg" alt=""></a></figure>
</div>
</div>
<p>
<p>
Line 76: Line 76:
</p>
</p>
-
<h4>Day 11 : 17-07-2012</h4>
+
<h4>Day 16 : 24-07-2012</h4>
<hr>
<hr>
<p>
<p>
-
Day 2 of making new competent cells <a href="http://partsregistry.org/Help:Protocols/Competent_Cells" target="_blanck" class="more">Read Protocol</a>
+
                                              Presence of colonies on both the positive and the negative control , we suspect a resistance from the competant cells or a bad selection with the plates.
-
                                                 </br>
+
                                                 <br/>
-
                                                We stopped incubating our cells at an OD600nm of 0.31
+
                                              We ran a test to check the cells and the plates by puting to incubate overnight at 37°C.
-
                                                 </br>
+
                                                 <br/>
-
                                                In the end we made 192 tubes of 50μL competent cells.
+
                                              one ampicilin plate + ampicilin extempo + competent cells , one ampicilin plate + competent cells and one empty plate.                                              
</p>
</p>
-
<h4>Day 12 : 18-07-2012</h4>
+
<h4>Day 17 : 25-07-2012</h4>
<hr>
<hr>
<p>
<p>
-
The competence test gave good results with a good amount of colonies on each plates ( ≈ 300 ) and no colonies on the control plate.
+
The plates were responsible of the non selection of the transformants.
-
</br>
+
                                                <br/>
-
Today we will transform DNA for every biobrick left.
+
                                                We made a new stock of LB plates with Ampicilin , Chlorenphenicol or Kanamicin.
-
</br>
+
                                                <br/>
-
We will use our new competent cells.
+
                                                We transformed the biobricks from operon 1 an 3 again but plated them on our new stock.
-
</p>
+
                                                <br/>
-
<div class="img-box">
+
                                                Except for I.A put into pre-culture of 2mL LB and I.B as we used the transformant from Day 15.
-
<figure><a href="#"><img src="https://static.igem.org/mediawiki/2012/c/c5/Biobricks.jpg" alt=""></a></figure>
+
-
</div>
+
-
<p>
+
-
We will proceed by heat-shock as described in the partregistry.org protocol <a href="http://partsregistry.org/Help:Transformation_Protocol" target="_blanck" class="more">Read Protocol</a>
+
</p>
</p>
-
<h4>Day 13 : 19-07-2012</h4>
+
 +
<h4>Day 18 : 26-07-2012</h4>
<hr>
<hr>
<p>
<p>
-
                                                Today we picked single colonies and put the to grow in two 5 mL LB + Ampicilin 100μg/mL
+
Today we did a mini prep for each 5 pre culture tubes of I.A
                                                 </br>
                                                 </br>
-
                                                Except for 3.B who gave no colonies
+
We transformed the rest of the biobricks , IC/IIB/IIC(2)/IID/IIIB/IVA/IVC
                                                 </br>
                                                 </br>
-
                                                We put the tubes to incubate overnight at 37°C
+
We put five colonies of ID/IE/IIIA/IIIC/IIID/IIIE/5A into a 2mL LB pre-culture
</p>
</p>
-
<h4>Day 14 : 20-07-2012</h4>
+
<h4>Day 19 : 27-07-2012</h4>
<hr>
<hr>
<p>
<p>
-
                                                After 13 hours incubating we did a miniprep with the tubes. We got really poor results on the nanodrop for every biobrick (>3ng/μL). As we had really small cells pellets after the first centrifugation we suspect we didn't have enough cells in the beginning.
+
Today we did a miniprep with the colonies put into pre-culture overnight.
-
</p>
+
                                                </br>
 +
We put five colonies of IB/IC/IIB/IIC(2)/IID/IIIB/IVA/IVC into a 2mL LB pre-culture.
 +
</p>
</div>
</div>

Revision as of 14:06, 31 July 2012

Note Book - iGEM Bordeaux 2012

  • Biology

Day 15 : 23-07-2012

Today we decided to change some of the biobricks to reduce the number of assemblies

We made a transformation for The first operon biobricks with 2μL DNA.
We also made a Positive control with a well known plasmid and water as a negative control.
We followed the Partregistry protocol as usual.
And Plated 20μL and 200μL of the transformants on LB plates.
We put the plates to grow overnight at 37°C.

Day 16 : 24-07-2012

Presence of colonies on both the positive and the negative control , we suspect a resistance from the competant cells or a bad selection with the plates.
We ran a test to check the cells and the plates by puting to incubate overnight at 37°C.
one ampicilin plate + ampicilin extempo + competent cells , one ampicilin plate + competent cells and one empty plate.

Day 17 : 25-07-2012

The plates were responsible of the non selection of the transformants.
We made a new stock of LB plates with Ampicilin , Chlorenphenicol or Kanamicin.
We transformed the biobricks from operon 1 an 3 again but plated them on our new stock.
Except for I.A put into pre-culture of 2mL LB and I.B as we used the transformant from Day 15.

Day 18 : 26-07-2012

Today we did a mini prep for each 5 pre culture tubes of I.A
We transformed the rest of the biobricks , IC/IIB/IIC(2)/IID/IIIB/IVA/IVC
We put five colonies of ID/IE/IIIA/IIIC/IIID/IIIE/5A into a 2mL LB pre-culture

Day 19 : 27-07-2012

Today we did a miniprep with the colonies put into pre-culture overnight.
We put five colonies of IB/IC/IIB/IIC(2)/IID/IIIB/IVA/IVC into a 2mL LB pre-culture.

Partners

Project

Others

Retrieved from "http://2012.igem.org/Team:Bordeaux/Fourth"