Team:Chiba/Calendar-Home/21 August 2008

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20 August 2008 <|> 22 August 2008

Team:Input
(-->20/8)

Inoculated transformants for 12 hours in LB 2mL containing Ampicillin.


 * BBa_R0051(2007)
 * BBa_R0051(2006)
 * BBa_J06650(2007)
 * BBa_J06650(2006)
 * BBa_J22136(2007)
 * BBa_J22141(2007)

BBa_J22141:After inoculated for 11 hours, added 20&mu;l IPTG and cultured one hour.


 * UV irradiation test (plate phase)
 * two plasmids from(JW1908)
 * (Repressor)-Ptrc-LuxR-Plux-cI-colE1-Amp-
 * (Reporter)-PcI-GFP-p15a-Cm-

exposure. LB-Amp,Cm or LB-Amp, Cm, 100nMAHL for 12h at 37&deg;C.
 * 1) Inoculated (Reporter) culture from glycerol stock(-80&deg;C freezer) in 2mL LB-Ampicillin, Chloramphenicol medium and LB-Ampicillin, Chloramphenicol, 100nmAHL mediumfor 12h, at 37&deg;C.
 * 2) We diluted 105-fold, and plated 20ul of the resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other.
 * 3) We cultured for 12h at 37&deg;C.
 * 4) We iraddiated UV each plate. Wavelength:254nm,distance from the UV lamp to the cultures were 14cm.
 * 5) We performed the same operations on plates following 9 or 21h of UV
 * 1) Colonies from both plates were picked and cultured in 2mL of

resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other.
 * 1) We diluted 105-fold, and plated 20ul of the

result  GFP fluorescence was observed from plates without AHL.
 * Colony count

Team:Communication

 * (20/8)--->PCR
 * BBa_C0070(2007)
 * BBa_C0070(2006)
 * BBa_C0076(2007)
 * BBa_C0078(2007)
 * BBa_C0078(2006)


 * 95°C,5min -> ( 95°C,1min -> 52°C,1min -> 72°C,1min )･･･25cycles -> 72°C,10min -> 6°C


 * --->Gel Check


 * Transformation
 * competent cells : XL10G
 * BBa_S03154(2007)
 * BBa_S03154(2006)
 * BBa_I9026(2007)
 * BBa_I9026(2006)
 * BBa_I9030(2007)
 * BBa_I9030(2006)

--->(23/8)Digestion

Team:Output
Transformation
 * BBa_J32007(2007)-->no colony-->no colony(2 times)-->colony(3times)
 * BBa_B0034(2007)-->colony