Team:IPN-UNAM-Mexico/Notebook/September

We going to change clormphenicol cassete Bba_P1010 for this we take off the ccdB gene and do restriction with P & E then ligate with Bba_K266006. Take out DNA from the well and electroporated the plasmid into Top 10 competent E. coli cells and plate, left overnight in 37ºC.

22-September-2009

We get colonies our ligation and left to incubate overnight on the shaker for midiprep.

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+==='''''01-September-2009'''''===
+We do the transformation by head shock and plate.
+----
+----
+==='''''02-September-2009'''''===
+We haven't transformant cells.
+----
+----
+==='''''07-September-2009'''''===
+Meeting, we have to take new strategy for this ligation, J23100 does't work as backbone but as insert is so short.....the strategy is try again.
+==='''''08-September-2009'''''===
+We left overnight ligation 14ºC
+----
+----
+==='''''09-September-2009'''''===
+Transformation for head shock and plate.  Get incubation overnight in 37ºC.
+----
+----
+==='''''10-September-2009'''''===
+We  have only two colonies and have to analyze we picked out an get in LB amp agar liquid, then we put on the shacker in 37ºC.
+----
+----
+==='''''11-September-2009'''''===
+Do midi prep run out gel; it dosen't mark the correct weight.
+----
+----
+==='''''14-September-2009'''''===
+We going to use J23100 as backbone and Bba_K266003 as insert. (2385 bp)
+----
+----
+==='''''21-September-2009'''''===
+We going to change clormphenicol cassete Bba_P1010 for this we take off the ccdB gene  and  do restriction with P & E  then ligate with  Bba_K266006.
+Take out DNA from the well and electroporated the plasmid into Top 10 competent E. coli cells and plate, left overnight in 37ºC.
+----
+----
+==='''''22-September-2009'''''===
+We get colonies our ligation  and left to incubate overnight on the shaker for midiprep.
+----
+----
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