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Team:Chiba/Calendar-Home/22 August 2008
From 2008.igem.org
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
- [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)
1. Incubated cultures(from Glycerol Stocks) with 2mL of LB-Ampicillin Medium for 12 hours at 37 degrees.
2. 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. )
4. covered the plates with polyethylene wrap.
5. after irradiation(Sample:15min,4h,8h,12h)(Negative Control:0h,1h,4h,8h)
UV irradiation test(Liquid phase)(~24)
-Ptrc-LuxR-Plux-cI-colE1-Amp-,-PcI-GFP-p15a-Cm-(BW,2plasmid)
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.
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。
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
- [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)
- PGEX VENUS YFP(GST fusion)
