Team:UNICAMP-Brazil/Notebooks/September 25

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ColiGuard

Getting the visa

  • Today was a happy day to five of us! We went to the U.S. Embassy in Sao Paulo and we got our visas! :D

finO and finP - Still Trying to Confirm our Biobricks

  • We ran an agarose gel of yesterday's PCRs product.

  • We couldn't obtain even a single amplified fragment! =(

  • Why can the transformed cells grew in the media (that means they actually have the AMP resistence), but they haven't our inserts into it's plasmids?

  • Our advisors suggested us that, since we digested our plasmid vector with XbaI and SpeI (X sticky ends can come together with S sticky ends), it recircularized without the introduction of our inserts.

  • Therefore, we must perform the dephosphorylation of our digested vector. That may prevent it from recircularizing without the insert introduction.

Marcelo

YeastGuard

Biofusion vector - Electroelution

  • Considering that we need more vector to perform the ligation reactions, we electroeluted more vector from the ADH1 biobrick previously digested with XbaI and SpeI (Protocol 12).

Taís