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Team:Newcastle/Slide Preparation
From 2010.igem.org
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Slide Preparation
- Grow overnight culture in 5 ml of LB broth, with appropriate antibiotics if required, at 37°C.
- Dilute the overnight culture to a ratio of 1:100 (overnight culture:LB with appropriate antibiotic if required). Incubate at 37°C for 1 hour.
- Set up the following broths with different concentrations of IPTG in 5 ml of LB broth:
- 2mM IPTG
- 0.2mM IPTG
- 0.02mM IPTG
- Broth only
- Innoculate the broth containing the different concentrations of IPTG with the appropriate cultures and incubate at 37°C for two hours.
- Prepare the slides by pippeting 500 µl of 1.2% agarose. Gently place the coverslip over the slide.
- Wait for about 5 minutes for the agarose to harden. Remove the coverslip by gently sliding the coverslip horizontally from the slide. This is to ensure that agarose layer remains undisturbed.
- Transfer 200 µl of each sample to microfuge tubes and label accordingly. Add 0.5 µl of membrane dye to each sample.
- Place 0.5 µl of each sample in each well of the slide and label accordingly. Place a coverslip on top of the slide.
- The slide is now ready for microscopy.
   
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