Team:Chiba/Calendar-Home/22 August 2008

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21 August 2008 <|> 23 August 2008

Contents

Laboratory work

Team:Input

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


insert check:Digestion

  1. BBa_J06650(2006)
  2. BBa_J06650(2007)
  3. BBa_J22136(2007)
  4. BBa_J22141(2007)
  5. BBa_R0051(2006)
  6. 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 37°C for 12h.

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


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

Team:Communication

Transformation
competent cells : XL10G
(Locate & 2 Punch ~iyama ver.~)


--->(24/8)Mini prep

Team:Output

Mini prep


Transformation

  • PGEX VENUS YFP(GST fusion)