Team:Valencia/Notebook/October

October 1st

- We repeated the previous experiment:
      Four strains in LCCs with SP medium.
      1 % Galactose induction at the beginning. Initial O.D. 2
      270 rpm.
       Initial temperature 28 ºC.

October 2nd

- Team meeting: We talked about the biobricks and some experiments left.

October 7th

- We repeated the previous experiment:
      Four strains in LCCs with SP medium.
      1 % Galactose induction at the beginning. Initial O.D. 2
      270 rpm.
       Initial temperature 28 ºC.

October 13th

- Total DNA was extracted from our yeast strains
- UCP-1 was amplified by PCR using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix.

October 14th

- We performed an agarose gel electrophoresis in order to check our PCR, the results was: Intense amplicons, of the expected size, (about 1 kb) were produced for UCP-1, 175-deleted and 76 deleted.

October 17th

- Team meeting: We discussed several things that need to be finished, such as sending DNA, preparing presentation, poster, wiki...

October 20th

- Preparing vectors, competent cells were transformed with: pSB1AK3 and pUC18 plasmids.

October 23th

- Plasmids were extracted (High pure miniprep plasmid isolation kit ROCHE).

- Plasmids were digested with EcoRI and PstI

October 24th

- Ligating Biobricks into plasmids: Inserts were run into agarose gels and DNA bands excised with a clean scalpel. DNA was extracted from agarose blocks (ultra clean gel spin, DNA purification Kit, MO BIO laboratories). T4 Ligase was used to ligate inserts and vectors.

- Competent cells were transformed and resulting colonies (Amp LB) screened with Fw and Rv primers to confirm the presence of inserts.

OCtober 27th

- pSB1AK3 containing UCP-1, 175-deleted and 76-deleted were sent to the Registry.