Team:EPF-Lausanne/Notebook/Wet Lab

From 2009.igem.org

(Difference between revisions)
(Removing all content from page)
 
(138 intermediate revisions not shown)
Line 1: Line 1:
-
{{EPF-Lausanne09}}
 
-
<div CLASS="epfltrick">__TOC__
 
-
</div><div CLASS="epfl09">
 
-
=Wet Lab=
 
-
==06.07.09==
 
-
LOVTAP plasmid AND TrpR plasmid were transformed in competent E. Coli following received protocol, and grown overnight (see Lab book for more details).
 
-
<br>One problem: we actually don't know TrpR plasmid resistance, so we tried with three resistances available in the lab: Amp., Kana. and Chl.
 
-
LOVTAP is in a plasmid called pCal-n (see picture below):
 
-
 
-
[[Image: pCAL-n.jpg|500px|thumb|center|pCal-n plasmid]]
 
-
 
-
<br>Some comments on the plasmid:
 
-
<br>-CBP is a small peptide with which we could purify LOVTAP protein
 
-
<br>-Thrombin target is a nucleotidic sequence that can be recognized by thrombin a peptidase. This peptidase will cut CBP once LOVTAP is purified
 
-
 
-
==07.07.09==
 
-
We have to grow the 3 strains generously sent by [mailto:j.beatty@ubc.ca Tom Beatty]
 
-
 
-
The three strains are :
 
-
:*''R.Palustris'' CEA001 (wild type) ; should be grown on LB medium only
 
-
:*''R.Palustris'' BPHP1+ ; should be grown on LB with gentamycin (100 micrograms/ml)
 
-
:*''E.Coli'' DH10B (pBPH/hmu0) ; should be grown on LB with gentamycin (20 micorgrams/ml)
 
-
 
-
 
-
The transformed LOVTAP and TrpR worked well (N.B. the plasmid of TrpR is pUC19 so the antibiotic resistance is Amp -> see below)
 
-
 
-
 
-
 
-
[[Image: RTEmagicC_puc19_2.gif.gif|500px|thumb|center|pUC19 plasmid]]
 
-
 
-
 
-
We did the glycerol stock, located in the -80 fridge, first floor of the iGEM compartement.
 
-
 
-
Then, a miniprep was done with both cultures.
 
-
A LOVTAP plasmid aliquot was done, a TrpR plasmid aliquot was done, located in the -20 fridge, 2nd floor.
 
-
 
-
==08.07.09==
 
-
 
-
==09.07.09==
 
-
 
-
==10.07.09==
 
-
 
-
</div><div CLASS="epfl09bouchon"></div>
 

Latest revision as of 08:05, 28 July 2009