Team:UNICAMP-Brazil/Notebooks/September 27

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 intend 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 XbaI 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 near. So, we decided to just isolate both CeaB and CeiB in a plasmid backbone. We chose to isolate into pSBA3 plasmid. Thus, we digested the plasmid with the right appropriate enzymes (SpeI and XbaI). Immediately, we made the purification gel according to 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

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