Oct 21, 2021
Revival of S. elongatus from plates
- 1IISER Pune
- iGEM IISER Pune India 2021
Protocol Citation: Akashdutta 2021. Revival of S. elongatus from plates. protocols.io https://dx.doi.org/10.17504/protocols.io.bzbsp2ne
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: October 21, 2021
Last Modified: October 21, 2021
Protocol Integer ID: 54354
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.
Take a 50 mL autoclaved flask.
Note
Remove the foil on the flask for aeration purposes.
Take a 10 mL 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, 150 rpm, 37°C