*<p style=”text-align:justify;”>We did mini-prep of the YFP+END negative colonies and then digested them with ''Xba''I and ''Pst'' I. We confirmed two digestions by electrophoresis and purified the correct fragment from the agarose gel (protocol 7).</p>
+
*<p style=”text-align:justify;”>We did mini-prep of the YFP+END negative colonies and then digested them with ''Xba''I and ''Pst'' I. We confirmed two digestions by electrophoresis and purified the correct fragment from the agarose gel ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Purification_of_DNA_fragments_from_agarose_gels Protocol 7]).</p>
We purify from agarose gel the digestion that we let doing yesterday and prepare another digestion with XbaI, this digestion was purified and used in a new ligation of BBa B0015 BBa K112806.
The ligation was used in a transformation, is the third time we do the same ligation, we hope this time we have luck!!!
Ane and Marcos
YeastGuard
New strategy: pGEM
We did miniprep of pDLD-pGEM, pJEN1-pGEM and Lysozyme-pGEM. We digested these plasmids with EcoRI and SpeI to release the parts from the pGEM. All of the digestions worked so we ligated them with biofusion vector previously digested with EcoRI and SpeI. We transformed competent E. coli with the promoters+biofusion ligation reaction and plated in LB+Amp media. The lysozyme ligation reaction was performed late at night, so we left it O/N to transform tomorrow.
GEL
GEL
YFP+Terminator
We did mini-prep of the YFP+END negative colonies and then digested them with XbaI and Pst I. We confirmed two digestions by electrophoresis and purified the correct fragment from the agarose gel (Protocol 7).
GEL
We performed the ligation reaction of YFP+END in Adh1.
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