Microbiology Notes CMOP April 2007 Cruise

April 4-7 2007, Wecoma – Oregon coastal zone (lines NH, SH, LB, CM and CR)

Cruise leader: Murray Levine

Microbiology sampling teams:
Team 1: (8-12) Lydie Herfort & Bill Howe
Team 2: (12-4) Dan Murphy & Michael Wilkin
Team 3: (4-8) Carolyn Sheehan & Dan Abraham

17 Stations samples for microbiology:
NH1, NH5, NH15, NH25, NH55
SH100, SH60, SH30,
LB30, LB60, LB100
CM25, CM10,
CR4, CR15, CR30, CR40

Samples collected (see attached excel file for volumes filtered):
o DNA/RNA, Sterivex: 4 with RNAlater and 2 with DEB buffer). Stored at -40oC. Sterivex: 1 until 6 = surface / 7 until 12 = bottom. DEB = 1 ml = 1, 2, 7, 8 & RNAlater = 2 ml = 3, 4, 5, 6, 9, 10, 11, 12.
o FISH: 40ml fixed with 1.6 ml of 37% filtered Formalin, incubated at room temp for 1 hr and then filtered (10 ml and 1 ml) 0.2 µm pore-size polycarbonate filters (Millipore, 25 mm diameter) using 0.45 µm pore-size cellulose nitrate supporting filters, let to dry and then stored at -40 oC. Remaining fixed sample was stored also in dry ice. Filtered using Manifold from Tebo’s lab.
o Nutrients: 40 ml of Sterivex filtrate collected in 50 ml Falcon tube, stored at -20oC.
o POC & PON: filtered onto 25 mm GFF filters (ashed over night at 500 oC), placed in ashed aluminium foil and stored at -20oC. Filtered using Manifold from Tebo’s lab.
o SPM: collected onto pre-weighted 25 mm GFF filters (filtered using Manifold from Tebo’s lab) and placed back into Eppendorf. Stored at -20oC.
o Chlorophyll a: 25 mm GFF filter (filtered using Manifold from Tebo’s lab), placed into orange screw-cap tubes

Comments:
NH1 - Cast 1
Note: 1h 30 min incubation for FISH instead of 1 h
Careful DEB = 1 & 6 and 7 &12

SH30
Position slightly different from usually because there were lots of crab pods in the coastal water.

LB30
Extra FISH collected at chlorophyll maximum at this site. Incubated for 1 h at room temp and stored at -40 oC (no filtration)

CR15
DEB added to 8 & 9 (instead of 7 & 8) and Sterivex filled up with DEB (= 2ml) instead of 1 ml.

CM25
Note: filter SPM ‘surface b’ is not good