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

1
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)
• CLPPDNA
• 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)
• Deadline40C 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
-20C. 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