Team:Cambridge/Week 3/Lab book

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{| style="color:#1b2c8a;background-color:#3ae2e8;" cellpadding="3" cellspacing="0" border="1" bordercolor="#fff" width="62%" align="center"
 
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!align="center"|[[Team:Cambridge|Home]]
 
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!align="center"|[[Team:Cambridge/Team|Team]]
 
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!align="center"|[https://igem.org/Team.cgi?year=2012&team_name=Cambridge Official Team Profile]
 
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!align="center"|[[Team:Cambridge/Project|Project]]
 
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!align="center"|[[Team:Cambridge/Parts|Parts Submitted to the Registry]]
 
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!align="center"|[[Team:Cambridge/Modeling|Modeling]]
 
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!align="center"|[[Team:Cambridge/Notebook|Notebook]]
 
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!align="center"|[[Team:Cambridge/Safety|Safety]]
 
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!align="center"|[[Team:Cambridge/Attributions|Attributions]]
 
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!align="center"|[[Team:Cambridge/Sponsors|Sponsors]]
 
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|}
 
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{|align="center"
 
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!align="center"|[[Team:Cambridge/Week 3|Diary]]         
 
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!align="center"|[[Team:Cambridge/Week 3/Lab book|Lab book]]
 
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|}
 
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===Monday===
 
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'''Making ''bacillus'' competent'''
 
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*'''Bacillus salts''' *10 made up and autoclaved.
 
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*'''Medium A base''' *10 made up (glucose will be added tomorrow)
 
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*''bacillus'' strain 168 streaked out and grown on plate overnight.
 
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===Tuesday===
 
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'''Making ''bacillus'' competent'''
 
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*Filtered glucose added to sterile '''Medium A base'''.
 
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*Sterile aliquots of salts and medium apportioned and put in fridge (10*50ml each)
 
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*
 
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===Wednesday===
 
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'''Making ''bacillus'' competent'''
 
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[[File:Bsubgrowth.png|300px|right]]
 
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*Colonies of strain 168 removed from plate and added to three separate conical flasks, each with 48ml of '''Medium A''' inside. OD650 readings taken until enough cells were added such that absorbance was between 0.1 and 0.2.
 
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*Flasks placed inside a shaking incubator (200rpm) and samples taken every 20mins until growth had leveled off.
 
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*Growth curves, as well as t0 (at 170mins) plotted below:
 
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*90 mins after t0 (at 260 mins), cells removed from shaking incubator.
 
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*0.45ml of '''Medium B''' pre-warmed in Eppendorf tubes.
 
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*20 individual samples of 0.05ml taken from each growth flask and added to Eppendorfs.
 
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*Eppendorfs placed back in shaking incubator for 60 mins with lids off.
 
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*Eppendorfs centrifuged at ~13000 rpm for 10 mins, supernatant removed.
 
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*60% glycerol added (0.5ml/tube) and tubes vortexed.
 
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*All 60 tubes now frozen at -80 °C.
 
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===Thursday===
 
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Test transformation of frozen ''bacillus'' stocks
 
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*Two samples of ''bacillus'' defrosted from different batches.
 
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*
 
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===Friday===
 
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Transformation of ''bacillus'' and e.coli with lux genes from 2010
 
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Testing of the spectral properties of filters
 
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*Blue , Green and Red filters were tested in different wavelengths (400 nm - 600 nm) in a spectrophotometer.
 
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*Excellent absorbance/emmitance results given by the blue and red filters near 580nm and 490nm respectively. 580nm is the wavelength where intensity of light is to be measured to identify the presence of mOrange. 490nm is the wavelength where maximum light intensity is expected with or without the presence of mOrange.
 
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[[File:MOrange_expected_graphs.jpg|250px||thumb|left]]
 
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[[File:Filter_spectrum.png|250px||thumb|left]]
 

Latest revision as of 13:17, 20 July 2012