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
CeiB: Searching the right colony
We decided to search in the plate possible cells with the right DNA insert. We made 15 colony of each plate. Unfortunately we didn’t get again.
Ane
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).
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