Transcript Slide 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
3 Mosses and/or peat: 0-5 cm below
5 Peat: 12.5-17.5 cm below the surface
7 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
TUM-BO (Andreas)
PLFA
-80 g fwt
- deep freezing immediately
after sampling, shipping under
frozen conditions on dry ice
3x
Take moss (if
3 present) and the
old peat surface
Mix
underneath the
of
vegetation
3 parts
5
within
each
slice
for each 7 Only to be
analysed if 5 fails to
purpose
be a good old peat
background sample
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
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.
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
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
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