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

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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 ->



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