Team:Valencia/Notebook/August

From 2009.igem.org

(Difference between revisions)
Line 156: Line 156:
- iGEM meeting.
- iGEM meeting.
 +
 +
===August 4th===
 +
 +
Today we have had a yeast-team meeting. We suspect that our yeasts are not growing because they died during the trip between Barcelona and Valencia. We have decided to spread the yeasts we have left in YPD medium that had already been prepared, but we are skeptical about the results. If it doesn't work, Joaquin Arinyo and his teammates will send us more aequorin-transformed yeasts.
 +
We have also made 1 liter of newn solid SD lacking Leu medium.
 +
 +
===August 5th===
 +
 +
Today we have prepared the Coelenterazine stock dissolution. First we "put" N2 gas through a Methanol dissolution in order to substitute the O2, an inhibitor for the coelenterazine. After 5 or 10 minutes we took 200 microL and added them to 50 microg of Coelenterazine.
 +
 +
Then we have distribute them in eppendorfs of 5 microL each and kept them in the fridge at -20º.
 +
 +
For other side, we doubt if our yeast in the YPD medium have grown or not... We think we should wait until tomorrow... T-T... Like Cristina says, our yeasts are like Peter Pan: they don't want to grow up...
 +
 +
===August 6th===
 +
 +
Yeast have grown. YEaaaaaaaaaaaaahhhh!!! We have spread them into the SD solid medium lacking Leu that we did yesterday, we will wait until tomorrow for the results. Also we have prepared new SD liquid medium lacking Leu and once prepared we will also spread our super-yeasts in it. Tomorrow we will start with the protocol (we hope).
 +
 +
===August 7th===
 +
 +
Yeast have grown in SD medium lacking Leu (bothsolid and liquid) so we have started the protocol. The OD was low (0'18) but we have prepared an 1'8 OD dissolution of the yeasts by centrifugation and dilution with SD medium lacking Leu.
 +
Allright... well... we have mesured the "luminiscence" and we have seen nothing. We've made the experiment in an espectophotometer and in the electrophoresis gel photograph machine. Before the mesurements we checked the OD and it was 0'34 so very far from 1'8. Maybe that's the problem. We're not depressed anyway, on monday we will come back to demonstrate those yeasts what we can do. Valencia's iGEM way.
 +
 +
===August 10th===
 +
 +
Today, we've made a lot of things. Yeasts haven't grown after three days in SD lacking Leu liquid medium at 30º. We've found out that we were preparing the medium in the wrong way.
 +
 +
The right way is:
 +
 +
for 500ml
 +
 +
2'5 g (NH4)2SO4
 +
 +
1 g Yeast Synthetic Drop-Out Medium Supplement Without Leucine
 +
 +
10 g Glucose
 +
 +
7'5 g Agar (only for the solid medium)
 +
 +
We have also found out that the minimal volume for the spectophotometer is 400 microL so we will double all the quantities of the protocol.
 +
 +
===August 11th===
 +
 +
We repeated the protocol and we mesured in the espectrophotometer but we saw nothing. We think it's too few sensitive and we're going to ask for a fluorimeter. We also did the experiment in GelDoc but, again, we saw nothing.

Revision as of 10:47, 12 September 2009


Home Team Project Human Practices Notebook News

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
2021 22 23 24 25 26
2728 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
1112 13 14 15 16 17
1819 20 21 22 23 24
2526 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
1516 17  18 19 20 21
2223 24  25 26 27 28
2930  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
2021 22  23 24 25 26
2728 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
2425 26   27 28 29 30
311 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
2122 23  24 25 26 27
2829 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
1213 14  15 16 17 18
1920 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
 910 11  12 13 14 15
1617 18  19 20 21 22
23 24 25  26 27 28 29
30  1  2  3  4  5  6


August 3rd

- Yeasts haven't grown yet.

- iGEM meeting.

August 4th

Today we have had a yeast-team meeting. We suspect that our yeasts are not growing because they died during the trip between Barcelona and Valencia. We have decided to spread the yeasts we have left in YPD medium that had already been prepared, but we are skeptical about the results. If it doesn't work, Joaquin Arinyo and his teammates will send us more aequorin-transformed yeasts. We have also made 1 liter of newn solid SD lacking Leu medium.

August 5th

Today we have prepared the Coelenterazine stock dissolution. First we "put" N2 gas through a Methanol dissolution in order to substitute the O2, an inhibitor for the coelenterazine. After 5 or 10 minutes we took 200 microL and added them to 50 microg of Coelenterazine.

Then we have distribute them in eppendorfs of 5 microL each and kept them in the fridge at -20º.

For other side, we doubt if our yeast in the YPD medium have grown or not... We think we should wait until tomorrow... T-T... Like Cristina says, our yeasts are like Peter Pan: they don't want to grow up...

August 6th

Yeast have grown. YEaaaaaaaaaaaaahhhh!!! We have spread them into the SD solid medium lacking Leu that we did yesterday, we will wait until tomorrow for the results. Also we have prepared new SD liquid medium lacking Leu and once prepared we will also spread our super-yeasts in it. Tomorrow we will start with the protocol (we hope).

August 7th

Yeast have grown in SD medium lacking Leu (bothsolid and liquid) so we have started the protocol. The OD was low (0'18) but we have prepared an 1'8 OD dissolution of the yeasts by centrifugation and dilution with SD medium lacking Leu. Allright... well... we have mesured the "luminiscence" and we have seen nothing. We've made the experiment in an espectophotometer and in the electrophoresis gel photograph machine. Before the mesurements we checked the OD and it was 0'34 so very far from 1'8. Maybe that's the problem. We're not depressed anyway, on monday we will come back to demonstrate those yeasts what we can do. Valencia's iGEM way.

August 10th

Today, we've made a lot of things. Yeasts haven't grown after three days in SD lacking Leu liquid medium at 30º. We've found out that we were preparing the medium in the wrong way.

The right way is:

for 500ml

2'5 g (NH4)2SO4

1 g Yeast Synthetic Drop-Out Medium Supplement Without Leucine

10 g Glucose

7'5 g Agar (only for the solid medium)

We have also found out that the minimal volume for the spectophotometer is 400 microL so we will double all the quantities of the protocol.

August 11th

We repeated the protocol and we mesured in the espectrophotometer but we saw nothing. We think it's too few sensitive and we're going to ask for a fluorimeter. We also did the experiment in GelDoc but, again, we saw nothing.













Back to July    |    August    |    Go to September