*<p style=”text-align:justify;”>Today we performed mini-preps ([https://2009.igem.org/Team:UNICAMP-Brazil/Protocols/Mini-Prep Protocol 2]) to extract the plasmids from the selected colonies inoculated yesterday.</p>
Another Miniprep, another digestion, another ligation and once again, another transformation
Transformation of the BBa K112806 + BBa B0015 ligation
We did the miniprep of BBa B0015 and BBa K112806 according to Protocol 2
With the protduct of the miniprep we made the digestion of BBa B0015 with EcoRI and XbaI and with EcoRI and SpeI to BBa K112806. After 3 hours of digestion we purified BBa K112806 from agarose gel and BBa B0015 directly from the digestion reaction.
Using the purified digestions we did the ligation, Protocol 11, of BBa B0015 with BBa K112806 and to test the digestion of BBa B0015 we did a ligation only without BBa K112806, if colonies grow with this ligation it means BBa B0015 is recircularizing. We staterd to quantify our samples with the new lab's espectrophotometer.
We transformed E. coli with the ligation, we hope this time it works!!
Ane and Marcos
PY Promoter - Mini-prep
Today we performed mini-preps (Protocol 2) to extract the plasmids from the selected colonies inoculated yesterday.
Fabi and Léo
YeastGuard
New strategy: pGEM
We need more parts!! We performed PCR to obtain the following parts: End and JENorf again.
We inoculated JENorf-pGEM and Jen1promoter-pGEM transformed colonies in liquid media to perform the screening tomorrow.
Terminator
We did PCR from the DNA ressuspended from the kit plate corresponding to the "Terminator" biobrick and from the DNA extracted form Kluyveyromices lactis. We found the JENorf fragment, unfortunately we didn't find the terminator expected fragment.
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