meth_1.12 Sponge Sampling for Marine Microbes Coastal Project

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Citation:

AM Project ID: Legacy

General Notes:

  1. First time the site is visited, sponge species abundance will be measured over 10 random quadrants. This will be repeated as required during the 14 months observation period.
  2. Select only sponge individuals that appear healthy and document specimen with a photo.
  3. Three different individuals will be collected at each sampling event.
  4. Record the following parameter for each individual: size (approximate in cm); covered with detritus (yes/no); any contact with other benthic, sessile organisms (yes, no; if yes, specify); morphological anomalies (discolouration; outgrowth etc.); epiphytic growth (type and coverage)

Method:

  1. Cut sponge samples underwater into small pieces (~5 g) using sterile blades. For each sponge specimen (n=3) collect six pieces each covering the entire body of the sponge
  2. Transfer pieces into 50 mL Falcon tube. Label containers according to the instruction given in the spreadsheet “sampling_schedule”.
  3. On the surface after the dive, use sterile gloves to squeeze water out of three sponge, pieces per sponge, transfer to a fresh 15 mL Falcon tube and plunge into liquid nitrogen (for RNA extraction). Label tubes according to the instruction given in the spreadsheet “sampling_schedule” and add “R” at the end (for RNA extraction).
  4. On the surface, wash three pieces three times in sterile seawater (5 min), transfer to a fresh 15 mL Falcon tube and then plunge into liquid nitrogen (for amplicon sequencing). Label tubes according to the instruction given in the spreadsheet “sampling_schedule” and add “A” at the end (for amplicon analysis).