Team:Chiba/Calendar-Home/22 August 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)

[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2007)と[http://partsregistry.org/Part:BBa_R0051 BBa_R0051](2006)はsingleのみ、ほかはsingleとdoubleの両方。

混ぜ表

	double	simgle
dH₂O	14	25
10×BSA	7	10
10×NE	7	10
EcoRI	3.5	5
PstI	3.5	-

UV照射実験(液体編)(~24)
-Ptrc-LuxR-Plux-cI-colE1-Amp-,-PcI-GFP-p15a-Cm-(BW)の2plasmidを使用。
グリストをつついて2ml培養×2(LB-Amp,Cm)(LB-Amp,cm,AHL100nM)
37℃で、12h培養後、濁ったら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。
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10⁵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。

37℃で12h培養した後のコロニー数

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

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。

Team:Communication

Transformation

competent cells : XL10G

BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]

(Locate & 2 Punch ~iyama ver.~)

BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
BBa_J04500(2008) [http://partsregistry.org/Part:BBa_J04500] -> No colonies on plates

--->(24/8)Mini prep

Team:Output

Mini prep

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

Transformation

PGEX VENUS YFP(GST fusion)