From 2014.igem.org
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Revision as of 20:45, 12 October 2014
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2D detection of IPTG and HSL
Chip production
Cell preparation
- 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 .
Agar preparation
- 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.
Chip preparation
- 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.
Measurement of fluorescence
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