Team:UNIPV-Pavia/Notebook/Week3Oct

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= Week from October 12th, to October 18th, 2009 =
= Week from October 12th, to October 18th, 2009 =
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct">
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct#week_start">
  <img src="https://2009.igem.org/Special:FilePath?file=Left_arrow.png"/>
  <img src="https://2009.igem.org/Special:FilePath?file=Left_arrow.png"/>
</a>
</a>
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct">Previous Week</a>
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct#week_start">Previous Week</a>
</td>
</td>
<td align="right" width="50%">  
<td align="right" width="50%">  
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct">Next Week</a>
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct#week_start">Next Week</a>
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct">
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct#week_start">
  <img src="https://2009.igem.org/Special:FilePath?file=Right_arrow.png"/>
  <img src="https://2009.igem.org/Special:FilePath?file=Right_arrow.png"/>
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== <html><font class="dayw_style">October, 12th</font></html> ==
 +
*We repeated the digestion for F2620TOP10(S-P) and B5new2-3(X-P-ClaI).
 +
*Gel run/cut for the two plasmids: bands were good.
-
== <html><font class="dayw_style">October, 12th</font></html> ==
+
*Gel extraction for:
 +
**F2620TOP10(S-P)
 +
**B5new2-3(X-P-ClaI)
 +
**B3(X-P)
 +
**B4(X-P)
 +
**A19-1(S-P)
 +
 
 +
*Unfortunately we had a very low DNA yield for almost all of them: A19-1(S-P) and F2620(S-P) could not be used :'(
 +
 
 +
*We decided not to repeat A19-1 extraction (and then the A21 re-cloning) and to assay our A21 construct for ethanol production.
 +
 
 +
*F2620 DNA was over, so we inoculated 8 ul of F2620MIT1 glycerol stock in 7 ml of LB + Amp. We incubated this inoculum overnight (37°C, 220 rpm).
 +
 
 +
*We inoculated 12 of our assemblies in 3 ml of LB + Amp in order to prepare the DNA to send to iGEM HQ:
 +
**A1
 +
**A2
 +
**A3
 +
**A4
 +
**A5
 +
**A6
 +
**A7
 +
**A8pg
 +
**A9pg
 +
**A11-3
 +
**A12-2
 +
**A15-3
 +
*We ordered a gas chromatography for a sample of the supernatants (taken the previous day).
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== <html><font class="dayw_style">October, 13th</font></html> ==
== <html><font class="dayw_style">October, 13th</font></html> ==
 +
*Miniprep for the 12 inocula to be sent.
 +
 +
*Miniprep, cut S-P, gel run/cut/purification for F2620MIT1. This time we had a good yield;)
 +
 +
*Ligations:
 +
**B14 = F2620(S-P) + B3(X-P) in pSB1A2
 +
**B15 = F2620(S-P) + B4(X-P) in pSB1A2
 +
*We incubated them at 16°C overnight.
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== <html><font class="dayw_style">October, 14th</font></html> ==
== <html><font class="dayw_style">October, 14th</font></html> ==
 +
 +
*We transformed the two ligations (after 1:20 dilution) and incubated the plated bacteria overnight (37°C).
 +
 +
*We inoculated all the remaining assemblies (12) in order to send purified DNA to iGEM HQ (3 ml of LB + Amp).
 +
 +
*We inoculated 8 ul of B5new2-3, F2620TOP10 and B0015 glycerol stocks in 8 ml of LB + Amp + 2% glucose. We incubated them at 37°C, 220 rpm in the morning.
 +
*Team meeting
*Team meeting
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[[#top|Top]]
[[#top|Top]]
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== <html><font class="dayw_style">October, 15th</font></html> ==
== <html><font class="dayw_style">October, 15th</font></html> ==
 +
*We diluted 25 ul of A14S-3 in 3 ml of LB + Amp and in 3 ml of LB + Amp + 1mM IPTG. We incubated the two cultures at 37°C, 220 rpm for 6 hours. Then we measured OD600 and fluorescence, but unfortunately their GFP levels were the same.
 +
 +
*Miniprep for the remaining BioBricks.
 +
 +
*pH measurement for B5new2-3, F2620TOP10 and B0015 24 hour cultures. We diluted 1:100 these three cultures in LB + Amp + 10% glucose and aliquoted them in the microplate reader (anaerobic).
 +
 +
*We inoculated 4 colonies of B14 plate and 3 colonies of B15 plate in 5 ml of LB + Amp and incubated them at 37°C, 220 rpm overnight. Tomorrow they will be screened!
 +
 +
*We received gas chromatography results.
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<div align="right">
[[#top|Top]]
[[#top|Top]]
</div>
</div>
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== <html><font class="dayw_style">October, 16th</font></html> ==
== <html><font class="dayw_style">October, 16th</font></html> ==
 +
*Cloning
 +
*Fermentation experiment setup
 +
*Wiki updating
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[[#top|Top]]
[[#top|Top]]
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== <html><font class="dayw_style">October, 17th</font></html> ==
== <html><font class="dayw_style">October, 17th</font></html> ==
-
 
+
*Fermentation experiment
 +
*Wiki updating
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<div align="right">
[[#top|Top]]
[[#top|Top]]
</div>
</div>
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== <html><font class="dayw_style">October, 18th</font></html> ==
== <html><font class="dayw_style">October, 18th</font></html> ==
-
 
+
*Fermentation experiment
 +
*Wiki updating
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<tr>
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<td align="left" width="50%">
<td align="left" width="50%">
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct">
+
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct#week_start">
  <img src="https://2009.igem.org/Special:FilePath?file=Left_arrow.png"/>
  <img src="https://2009.igem.org/Special:FilePath?file=Left_arrow.png"/>
</a>
</a>
-
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct">Previous Week</a>
+
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week2Oct#week_start">Previous Week</a>
</td>
</td>
<td align="right" width="50%">  
<td align="right" width="50%">  
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<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct">Next Week</a>
+
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct#week_start">Next Week</a>
-
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct">
+
<a href="https://2009.igem.org/Team:UNIPV-Pavia/Notebook/Week4Oct#week_start">
  <img src="https://2009.igem.org/Special:FilePath?file=Right_arrow.png"/>
  <img src="https://2009.igem.org/Special:FilePath?file=Right_arrow.png"/>
</a>
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Latest revision as of 23:40, 21 October 2009

EthanolPVanimation.gif

December 2008
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March 2009
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April 2009
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May 2009
M T W T F S S
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25 26 27 28 29 30 31
June 2009
M T W T F S S
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29 30
July 2009
M T W T F S S
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13 14 15 16 17 18 19
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27 28 29 30 31
August 2009
M T W T F S S
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3 4 5 6 7 8 9
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24 25 26 27 28 29 30
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September 2009
M T W T F S S
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28 29 30
October 2009
M T W T F S S
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November 2009
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9 10 11 12 13 14 15
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30

Week from October 12th, to October 18th, 2009

Previous Week Next Week

October, 12th

  • We repeated the digestion for F2620TOP10(S-P) and B5new2-3(X-P-ClaI).
  • Gel run/cut for the two plasmids: bands were good.
  • Gel extraction for:
    • F2620TOP10(S-P)
    • B5new2-3(X-P-ClaI)
    • B3(X-P)
    • B4(X-P)
    • A19-1(S-P)
  • Unfortunately we had a very low DNA yield for almost all of them: A19-1(S-P) and F2620(S-P) could not be used :'(
  • We decided not to repeat A19-1 extraction (and then the A21 re-cloning) and to assay our A21 construct for ethanol production.
  • F2620 DNA was over, so we inoculated 8 ul of F2620MIT1 glycerol stock in 7 ml of LB + Amp. We incubated this inoculum overnight (37°C, 220 rpm).
  • We inoculated 12 of our assemblies in 3 ml of LB + Amp in order to prepare the DNA to send to iGEM HQ:
    • A1
    • A2
    • A3
    • A4
    • A5
    • A6
    • A7
    • A8pg
    • A9pg
    • A11-3
    • A12-2
    • A15-3
  • We ordered a gas chromatography for a sample of the supernatants (taken the previous day).

Top

October, 13th

  • Miniprep for the 12 inocula to be sent.
  • Miniprep, cut S-P, gel run/cut/purification for F2620MIT1. This time we had a good yield;)
  • Ligations:
    • B14 = F2620(S-P) + B3(X-P) in pSB1A2
    • B15 = F2620(S-P) + B4(X-P) in pSB1A2
  • We incubated them at 16°C overnight.

Top

October, 14th

  • We transformed the two ligations (after 1:20 dilution) and incubated the plated bacteria overnight (37°C).
  • We inoculated all the remaining assemblies (12) in order to send purified DNA to iGEM HQ (3 ml of LB + Amp).
  • We inoculated 8 ul of B5new2-3, F2620TOP10 and B0015 glycerol stocks in 8 ml of LB + Amp + 2% glucose. We incubated them at 37°C, 220 rpm in the morning.
  • Team meeting

Top

October, 15th

  • We diluted 25 ul of A14S-3 in 3 ml of LB + Amp and in 3 ml of LB + Amp + 1mM IPTG. We incubated the two cultures at 37°C, 220 rpm for 6 hours. Then we measured OD600 and fluorescence, but unfortunately their GFP levels were the same.
  • Miniprep for the remaining BioBricks.
  • pH measurement for B5new2-3, F2620TOP10 and B0015 24 hour cultures. We diluted 1:100 these three cultures in LB + Amp + 10% glucose and aliquoted them in the microplate reader (anaerobic).
  • We inoculated 4 colonies of B14 plate and 3 colonies of B15 plate in 5 ml of LB + Amp and incubated them at 37°C, 220 rpm overnight. Tomorrow they will be screened!
  • We received gas chromatography results.

Top

October, 16th

  • Cloning
  • Fermentation experiment setup
  • Wiki updating


Top

October, 17th

  • Fermentation experiment
  • Wiki updating

Top

October, 18th

  • Fermentation experiment
  • Wiki updating

Top

Previous Week Next Week
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