Cyanophage plaque purification
Plate cells as described in Plating Prochlorococcus and Synechococcus strains in top agarose for plaque assays .
When plaques begin showing up , pick them using the “coring” technique .
To follow the liquid cultures , you just need to check ( by eye ) every couple of days to see if any of them have lysed .
When you check them , give them a quick LOW - SPEED vortex ( < 4 ) to get the cells off the bottom of the tube ( particularly the Syn ) without spilling cells .
When you go to “harvest” the cyanophage ( after lysis ) , decant the tube into two 50 ml orange capped tubes .
Spin on the Beckman JA - 17 rotor , 10K ( ~ 13,800g ) , 17°C , 20 minutes .
Transfer the supernatant into two fresh pre - labeled acid - washed glass black - capped tubes ( label with : cyanophage name , date , “spun” , 2xplaque purified ) for storage .
Work asceptically in the hood with flame .
