TABLE 3

Recovery of nucleic acids from microorganisms collected on AAO (0.02-μm Anotop 25) filters

ControlaSamplebIDExtraction methodcVariable% recoveryd
DNARNA
LiquidPhageAMPFlush direction73 (10)
BacteriaBMPFlush direction96 (7)
IMPChemistry, 184 (8)73 (5)
JMPChemistry, 2102 (4)91 (4)
ProtistHMPVol, 1 ml89 (12)
HMPVol, 2 ml82 (14)
HMPVol, 4 ml72 (11)
Seawater, KBDMP% recovery117 (23)
GMPVol, 10 ml130 (52)
GMPVolume 50 ml68 (53)
GMPVol, 250 ml93 (30)
GMPVol, 500 ml113 (36)
GMPVol, 1 liter95 (33)
All liquid93 (24)82 (12)
Pelleted*ProtistCMPFlush direction144 (12)
Seawater, AWKMP, 60Chemistry + Lys91 (11)
KMP, 60 + LysChemistry + Lys92 (14)
KLB1, 60Chemistry + Lys110 (38)
KLB1, 60 + LysChemistry + Lys65 (19)
Pelleted**Seawater, KBEMP% recovery112 (7)134 (15)
FMPVol,10 ml132 (33)
FMPVol, 50 ml69 (49)
FMPVol, 250 ml95 (6)
FMPVol, 500 ml115 (7)
FMPVol, 1 liter97 (15)
All pelleted102 (19)134 (15)
Pelleted** + supernatantSeawater, KBEMP% recovery106 (7)
  • a Extraction efficiency controls consisted of direct extractions of microorganisms, including the liquid in which they were suspended (liquid) or after centrifuging to pellet primarily cells (*) or cells and viruses (**) as indicated (pelleted).

  • b For environmental samples, locations were Ala Wai Canal (AW) and Kāne‘ohe Bay (KB).

  • c The extraction methods refer to the standard MasterPure protocol with 15 min of proteinase K incubation and no lysozyme treatment (MP), unless otherwise indicated by 60 (60 min of proteinase K incubation), + Lys (lysozyme treatment), or LB1 (nonproprietary extraction method).

  • d Values are means (standard deviations) from triplicate assays, expressed as a percentage of control values.