"
Team:Chiba/Calendar-Home/22 August 2008
From 2008.igem.org
(→Team:Input) |
(→Team:Input) |
||
| Line 59: | Line 59: | ||
UV irradiation test | UV irradiation test | ||
| - | * two plasmids from | + | * two plasmids from |
o -Ptrc-LuxR-Plux-cI-colE1-Amp- | o -Ptrc-LuxR-Plux-cI-colE1-Amp- | ||
o -PcI-GFP-p15a-Cm-(JW1908) | o -PcI-GFP-p15a-Cm-(JW1908) | ||
Revision as of 00:46, 30 October 2008
21 August 2008 <|> 23 August 2008
Contents |
Laboratory work
Team:Input
Mini prep: Minipreped the cultures(<--21/8).
- BBa_R0051(2007)
- BBa_R0051(2006)
- BBa_J06650(2007)
- BBa_J06650(2006)
- BBa_J22136(2007)
- BBa_J22141(2007)
insert check:Digestion
- BBa_J06650(2006)
- BBa_J06650(2007)
- BBa_J22136(2007)
- BBa_J22141(2007)
- BBa_R0051(2006)
- 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
- BBa_J13002(2007)
- BBa_J13002(2006)
- BBa_J04500(2007)
- BBa_J04500(2006)
- (Locate & 2 Punch ~iyama ver.~)
- BBa_C0076(2008) -> No colonies on plates
- BBa_C0077(2008) -> No colonies on plates
- BBa_C0078(2008) -> No colonies on plates
- BBa_J04500(2008) -> No colonies on plates
--->(24/8)Mini prep
Team:Output
- BBa_J52008
- BBa_F2620
- BBa_T9002
- BBa_J33202
- PUC19
- PGEX VENUS YFP(GST fusion)
- PGEX VENUS YFP(GST fusion)
