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Water Sample Filtration 1) Sterilize the tweezers and scissors with 70% EtOH. Make sure the equipment is dry before continuing. **If your water sample is very dirty, filter it over the mesh screen alone to remove particulates. Pour the filtered flow through water into a clean container and rinse the filter flask. Re-assemble the filter with a new 1L flask. 2) Carefully remove a filter membrane from the packaging with a pair of sterile tweezers. 8μM = parasites, dinoflagelletes 0.45μM = bacteria 0.22μM = viral particles, CONSERVE samples 3) Place the filter disc on the mesh screen in the vacuum filter with the hash marks FACING UP and clamp the apparatus together. Make sure the pieces are tightly clamped to ensure a good vacuum seal, but leave a bit of space between the clamp and apparatus. 4) Add a little bit of the water sample to the filter to moisten it, then turn on the vacuum. 5) Apply 500mL – 1L of sample to the filter; slowly pour the sample, a little bit at a time, onto the center of the filter. Monitor the vacuum pump to ensure it does not over-heat. Allow it to rest between uses if a lot of filtration needs to be done. 6) Allow the vacuum to run for a few seconds after all the liquid has passed through the filter, but do not over dry the filter disc. 7) Gently remove the filter from the screen with tweezers and take it to the fume hood. Using the tweezers and scissors in the hood, trim away excess filter. Roll the filter with the tweezers, and cut it into small pieces. Add the pieces to a bead tube and proceed to the appropriate DNA extraction protocol 8) Wash the filter flask, rinse it with EtOH, and hang to dry. Dissemble the filter apparatus and rinse the components with EtOH.