Team:BYU Provo/Notebook/CholeraDetection/Springexp/Period2/Dailylog

5/13/13

-KK Today we ran our digested CRO insert on the low melt gel, and under the UV light we saw the band we were looking for. Clarisse excised the band and we performed a ligation reaction according to the protocol and using the vector that we had already run on a low-melt gel last Thursday. After incubating for 30 minutes at room temperature, we performed a transformation into DH5alpha. The selectable marker of pLAT is ampicillin.

-KP We ran our low melt gel with our Cro insert. After 45 minutes, we put the gel under UV light and cut out the insert. We then performed a ligation of our vector and cro insert. After 30 min, we did a transformation and inserted our newly ligated plasmid into DH5alpha.

5/14/13

-KK When we dropped by today two of our plates (out of four) were contaminated. The two contaminated plates, we noticed, were the two plates that were old. The other two plates had a few colonies, but the was not a significantly greater number of colonies in our plasmid + insert + E.Coli plate than there were in our control plasmid + E.Coli plate. Kelton went ahead and streaked the few colonies we had to singles. We can PCR-verify if any of them have the plasmid. Meanwhile I am going to redo the transformation today. I will plate 4 plates: one 50 microL test, one 100 microL test, one 50 microL control, and one 100 microL control.

5/15/13

- Completed 5.9 T7+ Liquid Culture Phage Concentration Test #2 by doing the spot test

- Performed 5.15 Titer Test on 5.3 T7 new Phage Stock to determine phage concentration and estimate dilution for applying mutagen

- Sorted LB plates made on May 8 and threw away the ones with obvious contamination

5/16/13

- Took plates from 5.15 out of incubation at around 4:00pm

5/17/13

- Determined that all LB plates from 5.8 had contamination

- Poured new LB plates

- Made x8 top agar

5/18/13

- Stacked up the LB plates made yesterday. No obvious sign of contamination seen.

- Threw away the 5.8 LB plates (the ones with contamination).

5/19/13

- Started two 5mL of E coli BL21 overnight

- Designed procedure for applying mutagen and selecting for T7

5/20/13

- Started T7 Mutagen Concentration Test