====Transformation of the BBa K112806 + BBa B0015 ligation====
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*<p style=”text-align:justify;”>We transformed the ligation made yesterday by eletroporation according to the [https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Electroporation Protocol 3]</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
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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