Rig Filtration of SPOT 0m Water
Soak all small rig components in 10% acid for a minimum of 30 minutes. 
Rinse all components thoroughly using MilliQ (hold disks on edges).
Place on rig once rinsed.
Rinse red tubes and large plastic Erlenmeyer flask with MilliQ.
Assemble pump to rig using red tubes and large plastic Erlenmeyer flask.
Attach tube to right side of pump for vacuum.
Right knob on pump controls vacuum, loosen all the way. 
Remove filter top component and place upside down on bench. 
Remove micropore forceps from ethanol and rinse well with MilliQ.
Hold in a new sterile Falcon tube during use. 
Using micropore forceps remove 0.8μm backing disk from between blue disks and place on each rig filter. 
Again using the micropore forceps remove 0.2μm filter disk from between blue disks and place carefully on top of each 0.8μm disk.
Make sure they are centered and do not have holes or rips.
Place top component back on to each filter and twist on.
Open each rig filter by turning orange knob to verticle position.
Using a serological pipet and sterile glass pipet tip remove desired amount from bucket and pipet directly onto disks.
There should be 6 filtrations/filters of 10ml and 1 each of 20ml, 30ml, 40ml, 50ml, 60ml and 100ml.
Once desired amount of sample water is placed on filter and knobs are vertical, turn on pump.
Adujst top right knob on pump so that the arrow on the top right guage is slightly above 0.
Allow vacuum to work until most of the water has passed through the filters.
Once all water has passed through a filter, turn orange knob closed to the horizontal position.
Once all filtration has finished and all knobs are horizontal, turn off pump.
Twist off top component of rig filter and place upside down on bench.
Using the micropore forceps carefully remove only the top 0.2μm filter disk and place on a labeled TCBS Agar plate.
Use a sweeping or dragging motion to place filter flat on plate.
Repeat Step 12 for all 6 filters onto separate plates. 
Repeat Steps 7-13 for remaining volumes of water.
The 0.8μm filter disks may remain on the filters, only new 0.2μm filter disks are needed each time.
Leave plates right side up at room temperature on bench for at least 10 minutes.
Then place upside down in drawer to grow for at least 24 hours.
