"
Team:Valencia/Notebook/October
From 2008.igem.org
(→October 13th) |
|||
| Line 173: | Line 173: | ||
=== October 13th === | === October 13th === | ||
| - | - | + | - Total DNA was extracted from our yeast strains <br> |
| - | - | + | - UCP-1 was amplified by PCR using oligonucleotides matching the sequence and bearing the appropriate Biobrick prefix and suffix. |
=== October 14th === | === October 14th === | ||
| - | - We performed an agarose gel electrophoresis in order to check our PCR. | + | - 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 === | === October 17th === | ||
- Team meeting: We discussed several things that need to be finished, such as sending DNA, preparing presentation, poster, wiki... | - 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. | ||
| + | |||
| + | |||
Revision as of 18:48, 29 October 2008
|
|
|
| |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
|
|
|
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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.
