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Team:Chiba/Calendar-Home/5 September 2008
From 2008.igem.org
4 September 2008 <|> 6 September 2008
Contents |
Laboratory work
Team:Input
no work
Team:Communication
- (4/9)--->Liquid Culture
- Cultured the following cells (2mL LB-Amp, at 37°C)
- from transformed plates:
- BBa_K084009(Plac+RBS+RhlI+LVA, Competent Cells : JW1908)(r1, r3~r7)
- BBa_K084010(Plac+RBS+RhlI, Competent Cells : JW1908)(c1,c2,c4~c8)
- from Glycerol Stock:
- BBa_S03623(Ptet+RBS+LuxI, Competent Cells : JW1908)
- from transformed plates:
- --->Phenotype test
- MIX
Sample No. 1 2 3 4 5 6 7 8 9 10 11 r3 1mL - - - - - - - - - - r4 - 1mL - - - - - - - - - r6 - - 1mL - - - - - - - - r7 - - - 1mL - - - - - - - c1 - - - - 1mL - - - - - - c2 - - - - - 1mL - - - - - c6 - - - - - - 1mL - - - - c7 - - - - - - - 1mL - - - BBa_S03154[1] - - - - - - - - 1mL - - AHL(100μM) - - - - - - - - - - 1μL BBa_T9002[2] 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL IPTG(100μM) 1μL 1μL 1μL 1μL 1μL 1μL 1μL 1μL - - -
- Incubated for 8hours at 37 degrees
- The measurement of the intensity GFP by Fluoroskan Ascent 2.5(program:Ascent Software Version 2.6)
Sample No. 1 2 3 4 5 6 7 8 9 10 11 the intensity GFP (Frist run) 15.21 16.44 16.34 16.63 9.720 9.703 9.685 9.874 19.95 16.59 71.04 the intensity GFP (Second run) 17.85 19.19 19.28 19.14 9.981 10.16 10.14 10.41 23.94 17.61 79.53
- Spindown (max rpm, 3 min)
- The measurement of the intensity GFP(visual judgment) (UV 312nm)
Sample No. 1 2 3 4 5 6 7 8 9 10 11 the intensity GFP + + + + - - - - + ++ - Team:Output
DNA(digested sample) 2 PstI 0.1 BSA 0.8 Buffer3 1 dH2O 7 TOTAL 10μl -->37°C 30in
- BBa_J52008+BBa_F2620 x2(1)
- BBa_E2030+BBa_F2620 x2(2)
- BBa_J52008(PCR Product)(3)
- BBa_E2050(4)
Sample No. 1,2 3,4 DNA(digested sample) 10 1 loading Dye 2 1 TOTAL 12μl 6μl - Luciferase(BBa_J52008+BBa_F2620) check-->no light
