My lab has been having trouble making thick bacterial lawns. In a previous lab I routinely could put 2000 L1s on a 10cm seeded plate and they would still have food by the time they got to adulthood. Try as I might, I can’t seem to do that anymore. I’m not sure what the difference could be. Does anyone have any advice for getting thicker lawns?
Thanks much for any advice
Try increasing and playing around with the amount of peptone you add to your NGM recipe.
Good luck!
Strain NA22, which produces thicker lawns, can be substituted for OP50. It’s available from the stock center.
concentrate your bacteria 10x - 20x before seeding.
you could try to amplify single clone bacteria and put your plate with fresh lawn in 37C over night (or more hours). that always bring me thick lawns.
We always measure the OD(600) of each over-night OP50 LB culture and concentrate bacteria by centrifugation. With a concentrated pellet of 80 mL culture (OD~3) we cultivate up to 50.000 synchronized worms on one large dish.
We had issues with lawn thickness after switching peptone suppliers. We switched back to Difco peptone.