Week 9

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Revision as of 12:28, 12 September 2007 by Guido Costa (Talk | contribs)

  • Lab meeting on:


-LacI cloning under the pTetR R0040)promoter control.


  • There are colonies in the 07/30/07 plates.
  • We inoculate a E0020 and R0040 colony in 5ml.
  • We perform some tests to set up the fluorescence acquisition with the PMT (photomultiplier tube).

  • Miniprep of :

-I763012 + J06550

-I763012+ S0103 (I763016);

-B0034 + J22101 (missed in the sandbox);

-B0034 + J04631 (I763020);

-E0022 ;


  • Digestion for:

-I763012 + S0103 with Spe/Pst1;

-B0034 + J22101 with Xba/Pst1;

-B0034 + J04631 (I763020) with Xba/Pst1;

-E0022 with Xba/Pst1 ;

-R0040 with Spe/Pst1.

  • Band extraction from gel for:

-I763012+ S0103 ((I763016);

-B0034 + J22101 (missed in the sandbox);

-B0034 + J04631 (I763020);

-E0020 ;


  • Ligations at 15°C for 3h for:






  • Transformation of all ligations.
  • We find colonies for all ligations, except for the first ligation.
  • Photomultiplier adjustements

We decided to go on with our fluorescence measurements using not only the photo camera but also the photomultiplier described in the "Materials and Methods" section on our Homepage.

So, we have to adjust the PMT 814 features. First of all we calibrate this instrument by regulating the output offset to have a 0.00 output in case of photomultiplier in total darkness. Then, we adjust the PMT 814 time constant to 50 ns and its voltage gain to 10^(-3) microA/V. After that we