meth_1.9 Coastal Seawater Sampling for Downstream RNA Analyses for Marine Microbes Coastal Project

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AM Project ID: Legacy

An extension of SOP 1.17/18 Coastal Seawater Sampling for Downstream DNA Analyses, Marine Microbes Coastal Project:

  1. Use the same filtering set-up as in step 5 of SOP 1.17/1.18 to filter ~2 L of seawater separately three times onto three separate 0.22 micron sterivex filter. The filtering should be completed within 20 min of the water sample being collected.
  2. When the 2 L has been filtered continue to run the pump for 1-2 min to remove all of the liquid. If the filter gets blocked before, then note the approximate volume filter, disconnect the inlet and try to pump air through the filter for 1-2 min to remove remaining liquid.
  3. Ensure that the sterivex is labelled according to the instruction given in the spreadsheet “sampling_schedule” and append an “RNA” at the end.
  4. Cap the sterivex filter at one end and add 1.6 mL of RNAlater (Sigma). Cap the other end and shake well to distribute solution. Place the filter in a ziplock bag.
  5. Freeze immediately at -80 °C.