Team:UNICAMP-Brazil/Notebooks/October 6

ColiGuard

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

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.

GEL

Raíssa