For APA analysis, triplicate biological samples (250 mL) from in situ and incubation samples were filtered onto 47-mm polycarbonate membranes (0.2 μm). Stored at −20°C until analysis.
APA was assayed after Dyhrman and Ruttenberg (2006) using the fluorogenic phosphatase substrate 6,8-difluoro-4-methylumbelliferyl phosphate. Values were normalized to both volume and chl a. Reagents/Abs/Em used:
D-6567 6,8-difluoro-4-methylumbelliferyl phosphate (DiFMUP):
- Storage upon receipt: ≤ 20°C; Desiccate
- Abs/Em = 358/455
- Molecular Formula: C10H7F2O6P
- Molecular Weight: 292.1
- CAS Name/Number: 2H-1-Benzopyran-2-one, 6,8-difluoro-4-methyl-7-(phosphonooxy)-/ 214491-43-7
D-6566 6,8-difluoro-7-Hydroxy-4-Methylcoumarin (DiFMU) - Reference Standard:
- Storage upon receipt: Room temp.; protect from light
- Molecular Formula: C10H6F2O3
- Molecular Weight: 212.15
- CAS Name/Number: 2H-1-Benzopyran-2-one, 6,8-difluoro-7-hydroxy-4-methyl-/ 215868-23-8
Incubation key:
Control = no addition of nutrients or deep water
DSW = deep seawater addition (added 20% deep seawater (700 m))
+P = Added phosphate only (0.5 µM final for incubations 1 and 2, 1 µM final for incubation 3)
+N = Added nitrate only (6 µM final for incubations 1 and 2, 12 µM final for incubation 3)
phi_P = All but P added (N, Si, Fe, B12)
phi_N = All but N added (P, Si, Fe, B12)
-1, -2, -3 = biological replicates
In situ key:
IS = in situ
-1, -2, -3 = biological replicates
Lost = sample was lost