Team:Amsterdam/extra/protocols

From 2012.igem.org

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'''Growth curve time points'''<br\>
'''Growth curve time points'''<br\>
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The assessment Jose suggested seems fine.
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This contained a time laps of 30min for the first period of time (lets say first 3 hours) and after that time lapses of a hour would be better.
This contained a time laps of 30min for the first period of time (lets say first 3 hours) and after that time lapses of a hour would be better.
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*Prepare 2x 10ml pSB1AT3 + LacH cultures (in normal LB and with appropriate antibiotic added)
*Prepare 2x 10ml pSB1AT3 + LacH cultures (in normal LB and with appropriate antibiotic added)
*Prepare 2x 10ml pSB1AT3 + pBAD cultures (in normal LB and with appropriate antibiotic added)
*Prepare 2x 10ml pSB1AT3 + pBAD cultures (in normal LB and with appropriate antibiotic added)
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*Incubate at approximately 37 C for 20 hours (starting time 21:20)
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*Incubate at approximately 37 C for 20 hours  
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'''Day 2 (14-9-2012)'''<br\>
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'''Day 2 '''<br\>
Setup:
Setup:
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*Stop incubation of cultures after 20 hours of growth (at 16:20), cultures should now be well into the stationary faze.
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*Stop incubation of cultures after 20 hours of growth , cultures should now be well into the stationary faze.
*Prepare 8x a 500ml flasks containing 49.5ml of LB each
*Prepare 8x a 500ml flasks containing 49.5ml of LB each
*For 4 flasks add appropriate antibiotic for culture 1 (pSB1AT3 + LacH containing bacteria from day 1 - step 1)
*For 4 flasks add appropriate antibiotic for culture 1 (pSB1AT3 + LacH containing bacteria from day 1 - step 1)
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*To the 4 flasks from step 3 add 0.5ml of culture 1
*To the 4 flasks from step 3 add 0.5ml of culture 1
*To the 4 flasks from step 4 add 0.5ml of culture 2
*To the 4 flasks from step 4 add 0.5ml of culture 2
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*Incubate new cultures (step 5 & 6) at 37 degrees for 18 hours (starting time 18:00)
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*Incubate new cultures (step 5 & 6) at 37 degrees for 18 hours  
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'''Day 3 (15-9-2012)'''<br\>
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'''Day 3 '''<br\>
Experiment:
Experiment:
*Stop incubation of cultures after 18 hours of growth (at 12:00), cultures should now be well into the stationary faze.
*Stop incubation of cultures after 18 hours of growth (at 12:00), cultures should now be well into the stationary faze.

Revision as of 03:49, 27 September 2012