Team:Chiba/Calendar-Home/22 August 2008

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Colonies with no AHL plate observed GFP, 
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GFP fluorescence was observed from plates without AHL but not from
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いづれのプレートもAHLが入っていないものはGFPが確認できたが、AHL100nMのものは確認できなかった。
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plates containing 100nM AHL.
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Observe GFP fluorescence using UV exposure
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===Team:Communication===
===Team:Communication===

Revision as of 02:50, 30 October 2008

>Home | Notebook

21 August 2008 <|> 23 August 2008

Contents

Laboratory work

Team:Input

Mini prep: Minipreped the cultures(<--21/8).

  • [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)
  • [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
  • [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
  • [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
  • [http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
  • [http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)


insert check:Digestion

  1. [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2006)
  2. [http://partsregistry.org/Part:BBa_J06650 BBa_J06650](2007)
  3. [http://partsregistry.org/Part:BBa_J22136 BBa_J22136](2007)
  4. [http://partsregistry.org/Part:BBa_J22141 BBa_J22141](2007)
  5. [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)
  6. [http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)


Sample No. 1-41-6
dH2O(μl)1425
10×BSA(μl) 710
10×NE(μl) 710
EcoRI(μl) 3.55
PstI(μl) 3.5-
Total(μl) 3550


UV irradiation test

  • two plasmids from 
         o -Ptrc-LuxR-Plux-cI-colE1-Amp-
         o -PcI-GFP-p15a-Cm-(JW1908)
  1. Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin, Chloramphenicol Medium and LB-Ampicilline, Chloramphenicol and AHL100nm Medium for 12 hours at 37 degree.
  2. moved the cultures to small plates and started UV-irradiation. (wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)
  3. After exposing both plates to UV light for 15min, 4h, 8h, or 12h, we took 20ul from each plate, diluted 105-fold, and plated 20ul of the resulting solution t (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates.



The number of colonies aftetr culturing at 37C for 12h.

no AHLAHL100nM
15min 1360
4h 4031
8h 100
12h 00


GFP fluorescence was observed from plates without AHL but not from plates containing 100nM AHL.

Team:Communication

Transformation
competent cells : XL10G
  • [http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2007)
  • [http://partsregistry.org/Part:BBa_J13002　 BBa_J13002](2006)
  • [http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2007)
  • [http://partsregistry.org/Part:BBa_J04500　 BBa_J04500](2006)
(Locate & 2 Punch ~iyama ver.~)
  • [http://partsregistry.org/Part:BBa_C0076　 BBa_C0076](2008) -> No colonies on plates
  • [http://partsregistry.org/Part:BBa_C0077 BBa_C0077](2008)　-> No colonies on plates
  • [http://partsregistry.org/Part:BBa_C0078　 BBa_C0078](2008) -> No colonies on plates
  • [http://partsregistry.org/Part:BBa_J04500 BBa_J04500](2008) -> No colonies on plates


--->(24/8)Mini prep

Team:Output

Mini prep

  • [http://partsregistry.org/Part:BBa_J52008 BBa_J52008]
  • [http://partsregistry.org/Part:BBa_F2620 BBa_F2620]
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]
  • [http://partsregistry.org/Part:BBa_J33202 BBa_J33202]
  • PUC19
  • PGEX VENUS YFP(GST fusion)


Transformation

  • PGEX VENUS YFP(GST fusion)
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