Washed 4 day old QPX cultures. 3 isolates (S-1, TD-81, ATCC), two flasks (13mL) of each were washed in the following manner:
Each flask’s contents were transferred to a 50mL conical tube.
Each tube was topped with sterile sea water and mixed by inversion numerous times.
Tubes were centrifuged 10mins, 3000g at RT.
Transferred mucus and pellet to empty 50mL conical tube.
Topped with sterile sea water and mixed by inversion numerous times.
Centrifuged 10mins, 3000g at RT.
Removed as much supe/mucus as possible without disturbing the pellet.
Topped with sterile sea water and mixed by inversion numerous times.
Repeat steps 7 through 8 until sample is mucous-free and the pellet can be resuspended.
Pellets were eventually resuspended in 1mL of sterile sea water, split between two 1.5mL snap cap tubes and then brought up to 1mL with sterile sea water.