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

{| width="800"
 * style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]] Project name
 * style="background-color: #F2F2F2" align="center"|  |Main project page
 * style="background-color: #F2F2F2" align="center"|  |Main project page


 * colspan="2"|
 * colspan="2"|

{| width="800"
 * style="background-color: #cdde95;" align="center"|
 * style="background-color: #cdde95;" align="center"|




 * align="center" style="background-color: #e5edc8;" |

title=Search this Project


 * colspan="2" style="background-color: #F2F2F2;" align="right"|Customize your entry pages 
 * colspan="2"|
 * colspan="2"|
 * colspan="2"|

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 ->


 * }