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Week 6
7-13
Repeated conjugation procedure exactly as before except the strains are labeled H-P.
Repeated competent cell transformation with commercial competent cells (exactly the same as before).
7-14
Also, the conjugations all worked with the exception of one strain I had named 'N'. I plated them onto LBAmpKan to screen against all non conjugated bacteria and am letting them grow right now.
7-16
Finishing up conjugations, which did appear to grow. Frustratingly, the transformant did not grow this morning. I am giving it a little more time, but once more I feel as though the cells did not transform. I am beginning the procedure to make more competent cells for myself. I also may repeat the ligations and digests to be sure everything is ok.
7-17
I did a conjugation which grew on LB overnight, and then I plated it onto LBAmpKan and it did not grow, so I repeated the conjugation. This time it didn't grow on LB which doesn't really make sense unless there were no bacteria in the solution, which also doesn't make sense. The fact that none of them grew is concerning...I think maybe when I scraped the -80 bacteria I didn't get enough (or possibly any) bacteria. Will repeat conjugations on Monday, and prepare an overnight culture today.
