Team:Warsaw/Calendar-Stage1/6 July 2010

From 2010.igem.org

(Difference between revisions)
 
(8 intermediate revisions not shown)
Line 11: Line 11:
<!-- do not edit above me! -->
<!-- do not edit above me! -->
-
<div class="note"> Preparation of glycerol stocks</div>
+
<br> 6.07.2010 Tuesday
-
<br>1.Glycerol stocks of folloeing biobricks:  
+
<br>
 +
<br><div class="note"> Preparation of glycerol stocks [Dominik, Ania S., Kasia]</div>
 +
<br>1.Glycerol stocks of following biobricks:  
<a href="http://partsregistry.org/Part:BBa_J61100">J61100,</a>
<a href="http://partsregistry.org/Part:BBa_J61100">J61100,</a>
<a href="http://partsregistry.org/Part:BBa_J61101">J61101,</a>
<a href="http://partsregistry.org/Part:BBa_J61101">J61101,</a>
Line 26: Line 28:
[community RBSes],
[community RBSes],
<a href="http://partsregistry.org/Part:BBa_J23100">J23100</a>
<a href="http://partsregistry.org/Part:BBa_J23100">J23100</a>
-
J23100 [synthetic promoter].
+
[synthetic promoter].
<br>2. Alkaline lysis of part of liquid culture used to prepare stocks. Undigested plasmids run on the gel. Results seem to be correct.
<br>2. Alkaline lysis of part of liquid culture used to prepare stocks. Undigested plasmids run on the gel. Results seem to be correct.
<br>
<br>
-
<br><div class="note">Cloning of GFP-terminator behind RBSes - approach 2 </div>
 
-
<br>1. Digest GFP-terminator in psb1A2 plazmid. [16 μl DNA, 6 μl buforu, 16 μl H2O, 0,5 μl Xba/ Pst]
+
<br><div class="note">Cloning GFP-terminator behind RBSes - approach 2 [Ania P, Ania O., Kasia, Milena]</div>
-
<br>2. Gel extraction of Gel-out GFP-terminator.
+
 
-
<br>3. Ligation of GFPterminator with B0030, B0031, B0032, B0033, B0034 [community RBSes] [1 μl vector-RBSes, 10 μl < insert-GFPterminator, 2 μl buffer, 7 μl water]
+
<br>1. Digest of GFP-terminator in psb1A2 plasmid. [16 μl DNA, 6 μl buforu, 16 μl H2O, 0,5 μl Xba/ Pst]
 +
<br>2. GFP-terminator gel extraction.
 +
<br>3. GFPterminator ligation with  
 +
<a href="http://partsregistry.org/Part:BBa_B0030">B0030,</a>
 +
<a href="http://partsregistry.org/Part:BBa_B0031">B0031,</a>
 +
<a href="http://partsregistry.org/Part:BBa_B0032">B0032,</a>
 +
<a href="http://partsregistry.org/Part:BBa_B0033">B0033,</a>
 +
<a href="http://partsregistry.org/Part:BBa_B0034">B0034</a>
 +
[community RBSes],
 +
[1 μl vector-RBSes, 10 μl < insert-GFPterminator, 2 μl buffer, 7 μl water]
<br>4. Transformation with 20 μl ligation mixture
<br>4. Transformation with 20 μl ligation mixture
<br>
<br>
-
<br><div class="note">Cloning of GFP-terminator behind RBSes - approach 1 </div>
+
<br><div class="note">Cloning GFP-terminator behind RBSes - approach 1 [Ania S., Ania P., Kasia]</div>
-
<br>7. 05.07.2010 transformation results: contamination with ampicillin susceptible bacteria.
+
<br> 05.07.2010 transformation results: contamination with ampicillin susceptible bacteria.
-
<br>8. Set up one liquiquid culture fro each plate. Cultures inocculated with mix of all clones obtained on the plate.  
+
<br> Set up one liquid culture (3,5 ml) from each plate. Cultures inocculated with mix of all clones obtained on the plate.  
-
 
+
<br>
 +
<br>-> Repeated ligation of digested samples 2-6 (Anderson's RBSes) with gel-extracted GFP-terminator [Michał].
 +
<br>-> Transformation with the ligation mixtures.
 +
<br>
 +
<br>Summarised by Milena.

Latest revision as of 14:06, 31 July 2010

Example Tabs


6.07.2010 Tuesday

Preparation of glycerol stocks [Dominik, Ania S., Kasia]

1.Glycerol stocks of following biobricks: J61100, J61101, J61107, J61117, J61127 [Anderson's RBSes], B0030, B0031, B0032, B0033, B0034 [community RBSes], J23100 [synthetic promoter].
2. Alkaline lysis of part of liquid culture used to prepare stocks. Undigested plasmids run on the gel. Results seem to be correct.

Cloning GFP-terminator behind RBSes - approach 2 [Ania P, Ania O., Kasia, Milena]

1. Digest of GFP-terminator in psb1A2 plasmid. [16 μl DNA, 6 μl buforu, 16 μl H2O, 0,5 μl Xba/ Pst]
2. GFP-terminator gel extraction.
3. GFPterminator ligation with B0030, B0031, B0032, B0033, B0034 [community RBSes], [1 μl vector-RBSes, 10 μl < insert-GFPterminator, 2 μl buffer, 7 μl water]
4. Transformation with 20 μl ligation mixture

Cloning GFP-terminator behind RBSes - approach 1 [Ania S., Ania P., Kasia]

05.07.2010 transformation results: contamination with ampicillin susceptible bacteria.
Set up one liquid culture (3,5 ml) from each plate. Cultures inocculated with mix of all clones obtained on the plate.

-> Repeated ligation of digested samples 2-6 (Anderson's RBSes) with gel-extracted GFP-terminator [Michał].
-> Transformation with the ligation mixtures.

Summarised by Milena.

Retrieved from "http://2010.igem.org/Team:Warsaw/Calendar-Stage1/6_July_2010"