Team:TUDelft/13 July 2009

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='''13th July'''=
='''13th July'''=
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4 vials of DH5alpha comptetent cells were used (2 for each biobrick)
4 vials of DH5alpha comptetent cells were used (2 for each biobrick)
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after addition of 1 µl rehydrated biobrick DNA, the tubes were incubated for approx 15 (±5) min. on ice. Then one tube of each transformation was heatshocked for 1 min at 42 degrees.
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after addition of 1 µl rehydrated biobrick DNA, the tubes were incubated for approx 15 (±5) min. on ice. Then one tube of each transformation was heatshocked for 1 min at 42 degrees. No outgrowth phase was done, because amp resistance usually doesnt require this.
All four transformations were plated out and put at 37 degrees (at 19:00) <br>
All four transformations were plated out and put at 37 degrees (at 19:00) <br>
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BBa_K145280 heatshocked 50 microliter on agar+amp plate<br>
BBa_K145280 heatshocked 50 microliter on agar+amp plate<br>
BBa_K145279 heatshocked 50 microliter on agar+amp plate<br>
BBa_K145279 heatshocked 50 microliter on agar+amp plate<br>
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=='''Modelling'''==
 +
Worked on the delay device modelling (established the ODE's for cascade delay using formulas similar to the Bologna 2008 project)
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=='''Sriram'''==
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Prepared a plan for what to do in the lab next day. <br>
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Helped for modelling. <br>
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Checked whether all the biobricks are available. <br>
 +
Reviewed all the protocols that needs to be performed tomorrow. <br>
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Searched for some articles regarding conjugation modelling, and found two:
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*[http://www3.interscience.wiley.com/journal/118837812/abstract?CRETRY=1&SRETRY=0 A model for bacterial conjugal gene transfer on solid surfaces] and [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2K-4PT7X7G-1&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=fcacb99317376ced3df42e7e8178fc78 Rule-based modelling of conjugative plasmid transfer and incompatibility]
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=='''Calin'''==
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Ordered R751 containing cell culture (NCCBNr 2350) from Centraalbureau voor Schimmelcultures (http://www.cbs.knaw.nl).
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Latest revision as of 09:31, 26 July 2009

Lab Notebook

July
MTWTFSS
    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
August
MTWTFSS
          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
MTWTFSS
  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
MTWTFSS
      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
 

13th July

Weenink

prepared 10 agar+amp plates

Also did the first transformations to see how it goes:

BBa_K145280 (GFP-LVA + TetR generators)
BBa_K145279 (GFP + TetR generators)

4 vials of DH5alpha comptetent cells were used (2 for each biobrick)

after addition of 1 µl rehydrated biobrick DNA, the tubes were incubated for approx 15 (±5) min. on ice. Then one tube of each transformation was heatshocked for 1 min at 42 degrees. No outgrowth phase was done, because amp resistance usually doesnt require this.

All four transformations were plated out and put at 37 degrees (at 19:00)
BBa_K145280 no heatshock approx 80 microliter on agar+amp plate
BBa_K145279 no heatshock 50 microliter on agar+amp plate
BBa_K145280 heatshocked 50 microliter on agar+amp plate
BBa_K145279 heatshocked 50 microliter on agar+amp plate

Modelling

Worked on the delay device modelling (established the ODE's for cascade delay using formulas similar to the Bologna 2008 project)

Sriram

Prepared a plan for what to do in the lab next day.
Helped for modelling.
Checked whether all the biobricks are available.
Reviewed all the protocols that needs to be performed tomorrow.

Searched for some articles regarding conjugation modelling, and found two:

  • [http://www3.interscience.wiley.com/journal/118837812/abstract?CRETRY=1&SRETRY=0 A model for bacterial conjugal gene transfer on solid surfaces] and [http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6T2K-4PT7X7G-1&_user=10&_rdoc=1&_fmt=&_orig=search&_sort=d&_docanchor=&view=c&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=fcacb99317376ced3df42e7e8178fc78 Rule-based modelling of conjugative plasmid transfer and incompatibility]

Calin

Ordered R751 containing cell culture (NCCBNr 2350) from Centraalbureau voor Schimmelcultures (http://www.cbs.knaw.nl).