Team:UNICAMP-Brazil/Notebooks/September 27

ColiGuard

Inoculation of Yesterday's Ressuspended and Transformed Biobricks

• We selected 4 colonies from each plate containing one of yesterday's transformed biobricks, and inoculated them into liquid LB-AMP medium.

• Those inocula were grown for an O/N period at 37ºC (250 rpm).

Marcelo

Digestion of pTet and Double Terminator

• We pretend to ligate the parts which we are recovering right now (started yesterday) onto biobricks BBa_R0040 (pTet) and BBa_B0015 (double terminator), using it's own vector.

• Those biobricks were already recovered on August 10th.

• BBa_R0040 was digested with SpeI and PstI, once we intend to add a fragment in front of it (downstream). As for BBa_B0015, the digestion was realized with EcoRI and Xba restriction enzymes, once we need to insert a fragment behind it's part (upstream).

• Digestion lasted 3 hours.

Marcelo

CeaB and CeiB

• We don’t have time, the deadline is next. So, we decided just isolate both CeaB and CeiB in a plasmid backbone. We chose isolating into [http://partsregistry.org/Part:pSB1A3 pSBA3] plasmid. Thus, we digested the plasmid with the right restriction enzymes (SpeI and XbaI). Immediately, we made the purification gel according to the protocol 7.

Luige

Cre-Recombinase - New PCR

• After several attempts on correctly digesting Cre-Recombinase's fragment, we ended up without any sample! =/
• Therefore, today we repeated the PCR reaction for amplifying Cre-Recombinase without ATG, in order to generate more sample.

Víctor

YeastGuard

Dephosphorylation - CIAP test

• The E.coli didn´t grow in any of the plates. =[ We think that we didn´t plate an enough volume of bacteria. So we decided to replate a larger volume. We hope to find colonies tomorrow!

• Meanwhile we kept screening the old transformations. No sucsess. =(

Raíssa and Taís