Team:Aachen/Notebook/Protocols
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# Over night culture of sensor cells (50 mL in a 250 mL flask with) max. 16 h | # 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. | + | # Centrifuge all 50 mL by 3000 g for 10 min at RT (21 °C). |
# Discard the supernatant | # Discard the supernatant | ||
- | # Re-suspend the pellet in 1 mL medium | + | # Re-suspend the pellet in 1 mL tempered(~21 °C) LB-medium . |
'''Agar preparation''' | '''Agar preparation''' | ||
- | # Autoclave 50 mL medium with 1.5 % agarose | + | # 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 | + | # Cool it down to 45 °C in a water bath. |
'''Chip production''' | '''Chip production''' | ||
- | # Mix the cooled medium with the cells by inverting gently | + | # Mix the cooled medium with the cells by inverting gently. |
- | # Pour it in the chip form, avoiding bubble formation (!) | + | # Pour it in the chip form, avoiding bubble formation (!). |
- | # Wait for ca 20 min until the agar has solidified | + | # Wait for ca 20 min until the agar has solidified. |
- | # Cut out the chips with a scalpel | + | # Cut out the chips with a scalpel. |
- | # Put | + | # Put two chips into a labeled petri dish and store additional 4 chips in labeled petri dishs in the refrigerator. |
- | # Incubate for 1 h at 37 °C | + | # Incubate two chips for 1 h at 37 °C prior to induction. |
= Transformation = | = Transformation = |
Revision as of 16:09, 6 October 2014
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