Team:Valencia/Notebook/October

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April
Mo Tu We Th Fr Sa Su
31  1   2  3  4  5  6
 7  8   9 10 11 12 13
14 15  16 17 18 19 20
21 22  23 24 25 26 27
28 29  30  1  2  3  4
May
Mo Tu We Th Fr Sa Su
28 29  30  1  2  3  4
 5  6   7  8  9 10 11
12 13  14 15 16 17 18
19 20  21 22 23 24 25
26 27  28 29 30 31 1
June
Mo Tu We Th Fr Sa Su
26 27  28  29 30 31  1
 2  3   4  5  6  7  8
 9 10  11 12 13 14 15
16 17  18 19 20 21 22
23 24  25 26 27 28 29
30  1   2  3  4  5  6
July
Mo Tu We Th Fr Sa Su
30  1  2  3  4  5  6
 7  8   9 10 11 12 13
14 15  16 17 18 19 20
21 22  23 24 25 26 27
28 29  30 31   1  2  3
August
Mo Tu We Th Fr Sa Su
28 29  30 31  1  2  3
 4  5   6  7  8  9 10
11 12  13 14 15 16 17
18 19  20 21 22 23 24
25 26  27 28 29 30 31
September
Mo Tu We Th Fr Sa Su
 1  2   3  4  5  6  7
 8  9  10 11 12 13 14
15 16  17 18 19 20 21
22 23  24 25 26 27 28
29 30  1 2 3 4 5
October
Mo Tu We Th Fr Sa Su
29 30    1   2  3  4  5
 6  7   8 9 10 11 12
13 14  15 16 17 18 19
20 21  22 23 24 25 26
27 28  29 30 31  1  2
November
Mo Tu We Th Fr Sa Su
27 28  29 30 31  1  2
 3  4   5  6  7  8  9
10 11  12 13 14 15 16
17 18  19 20 21 22 23
24 25  26 27 28 29 30


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.



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