Week 11

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Revision as of 12:11, 12 September 2007 by Guido Costa (Talk | contribs)
  • Miniprep for







Digestion for:

-I763027 with Xba/Spe;

-I763028 with Spe/Pst1 and with Xba/Spe;

-I763019 with Xba/Spe;

-I763021+P0412 with Eco/Spe;

-I763021+S03520 with Eco/Spe;

-I763021+S0100 with Eco/Spe;

  • Band extraction from gel for all digestion;
  • We have problems with J52034 and with I763019.
  • Ligations for:

-I763028 + I763007

-S0100 + J04431, because we want to understand if LacI operates well;

-J06550 + J04631, with this ligation we want to understand if it operates well, because it doesn't leak.

  • Digestion for:

-I763020 with Xba/Pst1;

-J22101 with Xba/Pst1;

  • Testing our devices

We take 5 ml of LB culture ground in which we put a colony of bacteria which contain Ptet-LacI Plac-cI-LacY-GFP plasmid. We examine a drop of it using our fluorescence microscopy setup. In these conditions, the bacteria don’t beam fluorescence. Starting from this moment, we put into the culture ground 1 mM of IPTG every 30 minutes until it is 4 millimolar, checking each time any eventual fluorescence, with negative results. After diluting 1 ml of the previous one with 4 ml of LB, we add IPTG until 1 ml of our new solution is 10 mM, then 20 mM and finally 40 mM. Despite that, no fluorescence is visible. Every time we checked for fluorescence, we used not only the photo camera but also the photomultiplier; every measurement showed a 0.16-0.18 volts offset, despite the bacteria being non-fluorescent: at least, we regulated that offset!