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Team:Chiba/Calendar-Home/21 August 2008
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**(Reporter)-PcI-GFP-p15a-Cm- | **(Reporter)-PcI-GFP-p15a-Cm- | ||
| - | #Inoculated (Reporter) culture from glycerol stock(-80°C freezer) in 2mL LB-Ampicillin, Chloramphenicol medium and LB-Ampicillin, Chloramphenicol, 100nmAHL mediumfor 12h, at 37& | + | #Inoculated (Reporter) culture from glycerol stock(-80°C freezer) in 2mL LB-Ampicillin, Chloramphenicol medium and LB-Ampicillin, Chloramphenicol, 100nmAHL mediumfor 12h, at 37°C. |
#We diluted 10<sup>5</sup>-fold, and plated 20ul of the resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other. | #We diluted 10<sup>5</sup>-fold, and plated 20ul of the resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other. | ||
#We cultured for 12h at 37°C. | #We cultured for 12h at 37°C. | ||
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</tr> | </tr> | ||
</table> | </table> | ||
| - | + | result | |
| - | + | <BR> | |
| - | + | GFP fluorescence was observed from plates without AHL | |
===Team:Communication=== | ===Team:Communication=== | ||
Revision as of 03:00, 30 October 2008
20 August 2008 <|> 22 August 2008
Contents |
Laboratory work
Team:Input
(-->20/8)
Inoculated transformants for 12 hours in LB 2mL containing Ampicillin.
- [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)
BBa_J22141:After inoculated for 11 hours, added 20μl 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-
- Inoculated (Reporter) culture from glycerol stock(-80°C freezer) in 2mL LB-Ampicillin, Chloramphenicol medium and LB-Ampicillin, Chloramphenicol, 100nmAHL mediumfor 12h, at 37°C.
- We diluted 105-fold, and plated 20ul of the resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other.
- We cultured for 12h at 37°C.
- We iraddiated UV each plate. Wavelength:254nm,distance from the UV lamp to the cultures were 14cm.
- We performed the same operations on plates following 9 or 21h of UV
exposure.
- Colonies from both plates were picked and cultured in 2mL of
LB-Amp,Cm or LB-Amp, Cm, 100nMAHL for 12h at 37°C.
- We diluted 105-fold, and plated 20ul of the
resulting solution to (LB-Amp,Cm)(LB-Amp,cm,AHL100nM) plates each other.
- Colony count
| no AHL | AHL100nM | |
| 9h | 606 | 453 |
| 12h | 146 | 151 |
result
GFP fluorescence was observed from plates without AHL
Team:Communication
- (20/8)--->PCR
- [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2007)
- [http://partsregistry.org/Part:BBa_C0070 BBa_C0070](2006)
- [http://partsregistry.org/Part:BBa_C0076 BBa_C0076](2007)
- [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2007)
- [http://partsregistry.org/Part:BBa_C0078 BBa_C0078](2006)
| DNA Template | 1 |
| dNTP mix | 10 |
| Foward Primer | 5 |
| Reverse Primer | 5 |
| DNA polymerase TAQ | 1 |
| Thermopol Buffer | 5 |
| dH2O | 28 |
| TOTAL | 50μL |
- 95°C,5min -> ( 95°C,1min -> 52°C,1min -> 72°C,1min )･･･25cycles -> 72°C,10min -> 6°C
- --->Gel Check
|
|
- Transformation
- competent cells : XL10G
- [http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
- [http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2006)
- [http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
- [http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2006)
- [http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2007)
- [http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)
--->(23/8)Digestion
Team:Output
- [http://partsregistry.org/Part:BBa_J32007 BBa_J32007](2007)-->no colony-->no colony(2 times)-->colony(3times)
- [http://partsregistry.org/Part:BBa_B0034 BBa_B0034](2007)-->colony
