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WP2: How to sample in the field and dispatch ? (after Scheyern, vers. 26.05.05 )

5. 3. 7. 3. WP2: How to sample in the field and dispatch ? (after Scheyern, vers. 26.05.05 ). First, for bulk density: vol. of slice (known) + weight of slice + dry weight 105 0 C of subsample, also note: depth of roots & zone of discoloration on the PVC pots. UFZ (Antonis) DNA (protists)

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WP2: How to sample in the field and dispatch ? (after Scheyern, vers. 26.05.05 )

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  1. 5 3 7 3 WP2: How to sample in the field and dispatch ? (after Scheyern, vers. 26.05.05) First, for bulk density: vol. of slice (known) + weight of slice + dry weight 1050C of subsample, also note: depth of roots & zone of discoloration on the PVC pots • UFZ (Antonis) • DNA (protists) • Takes the DNA extractions of MLURI Mosses and/or peat: 0-5 cm below Peat: 12.5-17.5 cm below the surface WSL-AR FISH-bacteria (Andy) - 3 g fwt - Fixing in PFA, washed with PBS (see procedure next page) .(For moss layers, fix reverse of a 75 ml container) - Shipping in centrifugation tubes with screw cap in 70% EtOH. Take moss (if present) and the old peat surface underneath the vegetation Mix of 3 parts within each slice for each purpose 3 x Peat: 32.5-37.5 cm below the surface • MLURI (Rebekka) • CLPP+DNA • 8 g fwt for CLPP + 3 g fwt for DNA • Shipping of CLPP samples in PE bags with isolated ice packing wrapping (no direct contact to prevent freezing), deadline 40C during 14 days • Shipping of DNA samples immediately frozen at -20ºC on dry ice by next day delivery 5 Only to be analysed if 5 fails to be a good old peat background sample 7 ISTO (Fatima) C, N, S (LECO) +Monosach. + Amino acids 150 g fwt Shipping in small containers with isolated ice pack wrapping Deadline: 1-2 weeks at 40C TUM-BO (Andreas) PLFA -80 g fwt - deep freezing immediately after sampling, shipping under frozen conditions on dry ice • UFC-CE, AR-WSL (Daniel G) • Bacteria/protists etc. • 1 cm3 in center of slice, in small containers, fixing in 10 ml GA 2% final conc.(For moss layers, fix reverse of a 75 ml container) • Keep in dark • Shipping immediately with isolated ice pack wrapping • ECOBIO (Andre-Jean) • C/N in microbial biomass, mic. activity • 100 g fwt • Shipping in PE bags with isolated ice pack wrapping (no direct contact to prevent freezing) • Deadline:40C during 14 days

  2. Procedure for FISH samples for Andy (from Antonis) 4% Paraformaldehyde/PBS should be ok for FISH. I know that some flagellates (e.g. bodonids) will lyse during fixation with PBS, however at least for testate amoebae and bacteria this should be fine. Usually we fixe from 1-24h, wash twice with PBS and store in 70% ethanol at -20°C. Is there any problem with extended PFA-fixation (in case samples will be sent in PFA)? Does anybody need only ehtanol-fixed samples? Antonis For monosaccharides & amino acids, it would be preferable to take the old peat surface (slice of 5 cm thick) underneath the vegetation (same samples as those for Antonis). Is this the same for all people ? Fatima

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