Team:Chiba/Calendar-Home/22 August 2008

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UV irradiation test
UV irradiation test
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* two plasmids from ?
+
* two plasmids from  
           o -Ptrc-LuxR-Plux-cI-colE1-Amp-
           o -Ptrc-LuxR-Plux-cI-colE1-Amp-
           o -PcI-GFP-p15a-Cm-(JW1908)  
           o -PcI-GFP-p15a-Cm-(JW1908)  

Revision as of 00:46, 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)


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.
moved the cultures to small plates and started UV-irradiation. (wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)
after irradiation(Sample:15min,4h,8h,12h)(Negative Control(with AHL):0h,1h,4h,8h). After exposing both plates to UV light for 15min, 4h, 8h, or 12h, we took 20ul from each plate, diluted 10^5-fold, and plated 20ul of the resulting solution to (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


Colonies with no AHL plate observed GFP, 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。 Observe GFP fluorescence using UV exposure

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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