EPF-Lausanne/22 September 2008

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Sequencing

We got the sequence of AB and CD.

  • CD was just as expected.
  • There is an error with AB. The part B1 (RBS-LacIm) was not inserted correctly. We blasted the unknown sequence but were not able to find out where it came from. Anyways the test PCR before sequencing shows that it is probably an unwanted interaction, as it only worked when annealing temperature was reduced.

We have to redo the PCR of LacIm and the ligation with RBS.

Cloning

Agarose gel: Clones for the ligation RBS-LacIm
  • PCR of LacIm (reloaded)

We did a PCR on all plasmid samples (pHD) we had, mixed all results that were ok and inserted it into B0034.

Results:

One clone was correct.

RBS + LacIm = B1

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