*<p style=”text-align:justify;”>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 [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels protocol 7].</p>
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*<p style=”text-align:justify;”>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 [http://partsregistry.org/Part:pSB1A3 pSBA3] plasmid. Thus, we digested the plasmid with the right appropriate enzymes (''Spe''I and ''Xba''I). Immediately, we made the purification gel according to [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7].</p>
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 [http://partsregistry.org/Part:pSB1A3 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
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. =(
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