User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/07/05

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext">07/06/2010</div><div style="display:none;" id="page">User:Nelson Augusto Berrocal/Notebook/WiFi Coli 2010 Wet Lab/2010/07/05</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

Igem-logo-150px.png <sitesearch label="Search Lab Notebook" title="IGEM:HelpU/2008/Notebook/Project_name1">title=Search this Project</sitesearch>

Customize your entry pages Help.png

July 05th, 2010

LuxAB - J61002(containing constitutive promoter J23106) transformation resulted in two useful colonies out of 4 colonies; LuxAB - P18 transformation resulted in 1 useful colony; I recultured the three useful colonies (one LuxAB - P18, two LuxAB - J61002). Once recultures were grown enough I purified plasmid with High Pure Plasmid Purification Kit by Roche.
I extracted a total of 6 samples:
LuxAB - J61002 Colony 1 1
LuxAB - J61002 Colony 1 2

LuxAB - J61002 Colony 2 1
LuxAB - J61002 Colony 2 2

LuxAB - P18 1
LuxAB - P18 2

These are the order of the lanes of agarse gel 0.8% I ran to see if there were plasmids.

All 6 samples showed plasmids, now I'm gonna run a PCR to see if the insert of LuxAB is in the two vectors.
I ran a PCR with Taq Polymerase with 40 cycles:

5min 94°C --- | --- 45seg 94°C --- 45seg 55°C --- 3min 72°C (40 cycles) --- | --- 5min 72°C --- 4°C ->