meth_3.1.5 Qiagen DNeasy PowerSoil modified for seagrass Marine Microbes Coastal Projects

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AM Project ID: Legacy

  1. Extract each replicate swab (n=3) separately with the Qiagen DNeasy® PowerSoil® Kit.
  2. For this, aseptically transfer swab from cryotube to extraction tube by tipping from one to the other (or where necessary use clean, sterile forceps). Then follow the protocol as outlined in the manual, with the following modifications:
    • Step #5. Instead of using a vortex etc., replace this step by bead beating at full speed (30 Hz) for 10 min.
    • Step #18. Make sure filter is completely dry after this step. If not, repeat.
    • Steps #20-21. Split into 2 steps using 50 μL (rather than 100 μL once). Heat up solution C6 to 50 °C (when you start the whole lot), add 50 μL and leave it for 5 min, centrifuge at room temperature for 30 sec at 10,000 x g, and repeat (final volume = 100 μL).
  3. The final eluted DNA should be labelled according to the instruction given in the spreadsheet “sampling_schedule” and stored at -20 °C.