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Team:Valencia/Notebook/October
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<!-- ESCRIBIR AQUI --> | <!-- ESCRIBIR AQUI --> | ||
| + | |||
| + | === October 1st === | ||
| + | - We repeated the previous experiment:<br> | ||
| + | Four strains in LCCs with [[Team:Valencia/Notebook/Protocols#SP liquid medium | SP medium]]. <br> | ||
| + | 1 % Galactose induction at the beginning. Initial O.D. 2 <br> | ||
| + | 270 rpm. <br> | ||
| + | Initial temperature 28 ºC.<br> | ||
| + | |||
| + | === October 2nd === | ||
| + | - Team meeting: We talked about the biobricks and some experiments left. | ||
| + | |||
| + | === October 7th === | ||
| + | - We repeated the previous experiment:<br> | ||
| + | Four strains in LCCs with [[Team:Valencia/Notebook/Protocols#SP liquid medium | SP medium]]. <br> | ||
| + | 1 % Galactose induction at the beginning. Initial O.D. 2 <br> | ||
| + | 270 rpm. <br> | ||
| + | Initial temperature 28 ºC.<br> | ||
| + | |||
| + | === October 13th === | ||
| + | - Total DNA was extracted from our yeast strains <br> | ||
| + | - UCP-1 was amplified by [[Team:Valencia/Parts#Preparing_inserts|PCR]] using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix. | ||
| + | |||
| + | === October 14th === | ||
| + | - We performed an [[Team:Valencia/Parts#Preparing_inserts|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 === | ||
| + | - [[Team:Valencia/Parts#Ligating_Biobricks_into_plasmids|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. | ||
Latest revision as of 22:14, 29 October 2008
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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.
