Team:UNICAMP-Brazil/Notebooks/October 14

From 2009.igem.org

(Difference between revisions)
(→PY Promoter - Preparation of electrocompetent cells of the conjugative strain)
(→New strategy: pGEM)
Line 15: Line 15:
==''' YeastGuard '''==
==''' YeastGuard '''==
-
====New strategy: pGEM====
+
====[https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/pGEMStrategy New Strategy: pGEM]====
*<p style=”text-align:justify;”>We did miniprep of 10 Adh1+Lysozyme colonies.</p>
*<p style=”text-align:justify;”>We did miniprep of 10 Adh1+Lysozyme colonies.</p>

Revision as of 21:43, 21 October 2009

Topo l2.gif topo_r_igem.gif
topo_r_b.gif
Back to Calendar

ColiGuard

PY Promoter - Preparation of electrocompetent cells of the conjugative strain

  • Now that we have PY1 promoter + RFP inserted in a plasmid with kanamycin resistance we are able to transform the conjugative strain. As we said before, this conjugative strain already has ampicilin resistance, which is why we inserted our construction in a plasmid with a different resistance.

  • Before transforming this strain we need to prepair electrocompetent cells. So today we inoculated a colony of this strain in liquid LB-AMP at 37ºC to prepare the competent cells tomorrow.

Fabi and Léo

YeastGuard

New Strategy: pGEM

  • We did miniprep of 10 Adh1+Lysozyme colonies.

  • We digested the Adh1+Lysozyme/biofusion minipreps in order to connect it to the yeast expression vector (YEP358). We used the XbaI and PstI enzymes. 9 of 10 digestions worked! =) The expected fragment size is 2000bp.

Adh1+lis.jpg

  • We chose two fragments and purified them from the agarose gel. We ligated them to YEP358. We will transform it tomorrow.


Raíssa and Taís




Retrieved from "http://2009.igem.org/Team:UNICAMP-Brazil/Notebooks/October_14"