*<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 pSB1A3 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>
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 pSB1A3 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
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!
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