Team:Aachen/Notebook/Protocols
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+ | = 2-dimensional detection of IPTG and HSL = | ||
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+ | == Chip production == | ||
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+ | === Cell preparation === | ||
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+ | # over night culture of sensor cells (50 mL in a 250 mL flask with) max. 16 h | ||
+ | # centrifuge all 50 mL by 3000 g for 10 min at RT (21 °C). | ||
+ | # discard the supernatant | ||
+ | # re-suspend the pellet in 1 mL tempered (~21 °C) LB-medium . | ||
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+ | === Agar preparation === | ||
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+ | # autoclave 50 mL medium with 1.5 % (w/v) agarose (has to be multiplied with the number of chips prepared). | ||
+ | # cool it down to 45 °C in a water bath. | ||
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+ | === Chip preparation === | ||
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+ | # mix the cooled medium with the cells by inverting gently. | ||
+ | # pour it in the chip form, avoiding bubble formation (!). | ||
+ | # wait for approximately 20 min until the agar has solidified. | ||
+ | # cut out the chips with a scalpel. | ||
+ | # put two chips into a labeled petri dish and store additional 4 chips in labeled petri dishs in the refrigerator. | ||
+ | # incubate two chips for 1 h at 37 °C prior to induction. | ||
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+ | == Measurement of fluorescence == | ||
= Culture medium and culture conditions = | = Culture medium and culture conditions = | ||
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# after autoclaving, add 20 mM sterile glucose solution (filter sterilization) | # after autoclaving, add 20 mM sterile glucose solution (filter sterilization) | ||
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= Genetic methods = | = Genetic methods = |
Revision as of 14:33, 8 October 2014
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