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Team:Valencia/Notebook/October
From 2008.igem.org
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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 23rd
- 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.
