# Team:Valencia/Notebook/September

(Difference between revisions)
 Revision as of 08:48, 15 September 2009 (view source)Angeles (Talk | contribs)← Older edit Revision as of 21:38, 16 September 2009 (view source)Guimar3 (Talk | contribs) Newer edit → Line 184: Line 184: 1 = wt (1 microlitre) 1 = wt (1 microlitre) + 2 = wt (2 microlitres) 2 = wt (2 microlitres) + 3 = Cch1 (1 microlitre) 3 = Cch1 (1 microlitre) - 4 = Mid1 ('') + + 4 = Mid1 (1 microlitre) + 5 = Negative Control 5 = Negative Control + 6 = Possitive Control 6 = Possitive Control + (We used the same MWM) (We used the same MWM) Line 195: Line 201: We haven't obtain any result. We haven't obtain any result. - - - - - - Line 212: Line 212: + '''Programme''' - Programme After 3 minutes at 94 ºC as a previous step. Every of our 30 cycles has these trhee steps: After 3 minutes at 94 ºC as a previous step. Every of our 30 cycles has these trhee steps: - 30'' at 94 ºC - 1' at 55º - 1' at 72 ºC - We have added a final step of 7 minutes at 72ºC. + - 30s at 94ºC + - 1min at 55ºC + + - 1min at 72ºC + + We have added a final step of 7 minutes at 72ºC. 1 = wt (1 microlitre) 1 = wt (1 microlitre) + 2 = wt (2 microlitres) 2 = wt (2 microlitres) + 3 = Cch1 (1 microlitre) 3 = Cch1 (1 microlitre) - 4 = Mid1 ('') + + 4 = Mid1 (1 microlitre) + 5 = Negative Control 5 = Negative Control + 6 = Possitive Control 6 = Possitive Control + (We used the same MWM) (We used the same MWM) Wiiiiiiiiiiiiiiiii!!!!!!!!! We had amplification!!!!! We will use wt 1 microlitre of PCR amplification product (career 1) to build our first BioBrick in the next days. Wiiiiiiiiiiiiiiiii!!!!!!!!! We had amplification!!!!! We will use wt 1 microlitre of PCR amplification product (career 1) to build our first BioBrick in the next days. - - - - - Line 248: Line 251: ===15th September=== ===15th September=== + We have started with bacteria transformation. To build our first BioBrick, we selected the pSB1A3 plasmid located at 1K hole of plate 1 (distributed for the iGEM at spring 2009). We have used a E.coli strain ultracompetent called XL1-Gold. Transformation protocol comes with the strain. We have started with bacteria transformation. To build our first BioBrick, we selected the pSB1A3 plasmid located at 1K hole of plate 1 (distributed for the iGEM at spring 2009). We have used a E.coli strain ultracompetent called XL1-Gold. Transformation protocol comes with the strain. + After the steps, we left our strain in incubation, to make them add the plasmid. After the steps, we left our strain in incubation, to make them add the plasmid.

## Revision as of 21:38, 16 September 2009

 April Mo Tu We Th Fr Sa Su 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
 May Mo Tu We Th Fr Sa Su 27 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
 June 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
 July 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
 August 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 31 1 2 3 4 5 6
 September 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
 October 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
 November 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

### September 4th

We have tried our protocol using the fluoriscense microscope.We obtained some interesting results, but finally they were only artifacts. It seems like the the microscope is a good option to "see" our yeasts.

### September 11th

Today we have had an iGEM meeting.

We have repited the PCR with new primers:

Forward: 5'gaattcgcggccgcttctagatgaccagcgaccaatactc 3’

Reverse: 5’tactagtagcggccgctgcagttaggggacagctccaccg 3’

Programme After 2 minutes at 94 ºC as a previous step. Every of our 30 cycles has these trhee steps: 45 at 94 ºC 1' at 60º 1' at 72 ºC

1 = wt (1 microlitre)

2 = wt (2 microlitres)

3 = Cch1 (1 microlitre)

4 = Mid1 (1 microlitre)

5 = Negative Control

6 = Possitive Control

(We used the same MWM)

We haven't obtain any result.

### 14th September

Trying another PCR...

We have variated the programme another time T-T

Programme

After 3 minutes at 94 ºC as a previous step. Every of our 30 cycles has these trhee steps:

- 30s at 94ºC

- 1min at 55ºC

- 1min at 72ºC

We have added a final step of 7 minutes at 72ºC.

1 = wt (1 microlitre)

2 = wt (2 microlitres)

3 = Cch1 (1 microlitre)

4 = Mid1 (1 microlitre)

5 = Negative Control

6 = Possitive Control

(We used the same MWM)

Wiiiiiiiiiiiiiiiii!!!!!!!!! We had amplification!!!!! We will use wt 1 microlitre of PCR amplification product (career 1) to build our first BioBrick in the next days.

For other side, we have prepared Arinyo's protocol only with our wt strain to characterize the behaviour of our yeast with different amounts of KOH as an input. We didn't obtain any result.

### 15th September

We have started with bacteria transformation. To build our first BioBrick, we selected the pSB1A3 plasmid located at 1K hole of plate 1 (distributed for the iGEM at spring 2009). We have used a E.coli strain ultracompetent called XL1-Gold. Transformation protocol comes with the strain.

After the steps, we left our strain in incubation, to make them add the plasmid.

For other side, we have prepared Arinyo's protocol only with our wt strain to characterize the behaviour of our yeast with different amounts of KOH as an input.