Team:Chiba/Calendar-Home/22 August 2008

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Revision as of 00:45, 30 October 2008

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21 August 2008 <|> 23 August 2008

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

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

Sample No.	1-4	1-6
dH₂O(μl)	14	25
10×BSA(μl)	7	10
10×NE(μl)	7	10
EcoRI(μl)	3.5	5
PstI(μl)	3.5	-
Total(μl)	35	50

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 AHL	AHL100nM
15min	136	0
4h	40	31
8h	10	0
12h	0	0

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)

@@ Line 62: / Line 62: @@
            o -Ptrc-LuxR-Plux-cI-colE1-Amp-
            o -PcI-GFP-p15a-Cm-(JW1908)
-. Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
-<BR>
-. moved the cultures to small plates and started UV-irradiation.
-(wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)(Put the Negative control plate ina dark place. )
-<BR>
-. covered the plates with polyethylene wrap.
-<BR>
-. after irradiation(Sample:15min,4h,8h,12h)(Negative Control:0h,1h,4h,8h)
-<BR>UV irradiation test(Liquid phase)(~24)
-<BR>-Ptrc-LuxR-Plux-cI-colE1-Amp-,-PcI-GFP-p15a-Cm-(BW,2plasmid)
 <BR>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.
-<BR>そのまま小さなプレートにLB培地ごと移してUVを照射する。UVの波長は254nm,UVランプまでの距離は8cm。
+<BR>moved the cultures to small plates and started UV-irradiation. (wavelength:254nm,distance from the UV lamp to the cultures were 8cm.)
-<BR>UVを照射して、15min,4h,8h,12h,たったら、両方のプレートからLB培地ごと20μlずつ採取しそれぞれ10<sup>5</sup>希釈し、そこから20μlをそれぞれ(LB-Amp,Cm)(LB-Amp,cm,AHL100nM)のプレートに撒いた。
+<BR>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.
 <BR>The number of colonies aftetr culturing at 37C for 12h.