Revival of S. elongatus from plates

Abstract

On receiving wild type or tranformed S. elongatus plates, one needs to revive them for further studies. Revival of S. elongatus involves making its liquid culture in BG-11 medium. The following protocol describes the liquid culture preparation from BG-11 plates.

Steps

Take a 50mL autoclaved flask.

Note

Remove the foil on the flask for aeration purposes.

Take a 10mL aliquot of BG-11 solution in a falcon tube and transfer it into the flask. Keep the flask aside

Note

Carry out this process and all the successive steps inside a laminar airflow hood

Take the cyanobacteria plate and gently scrape it with the tip of a 1 mL loaded micropipette.

Note

Position the plate such that the streaked part faces the roof of the laminar hood, remove the lid carefully and place it upside down.

Dip the pipette tip in the aliquot of the BG-11 medium and gently mix.

Note

To mix, gently resuspend the liquid 1-2 times.

For the engineered strains, apply appropriate antibiotics (10μg/mL kanamycin and 4μg/mL gentamicin) in the BG-11 medium.

Store all the flasks in a photo incubator at 15000 lux, 150rpm

Abstract

Steps

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