268 
Fishery Bulletin 11 6(3-4) 
Map of stations ( + ) off Oregon and Washington sampled for species 
of Sebastes during 2005-2008. Along transects at Heceta Head, New¬ 
port, the Columbia River, and Willapa Bay, 5 stations were sampled 
during most sampling cruises. In addition, a single station located 
185 km from shore on the Newport transect (the NH-100 station) 
was sampled once in June 2008. 
riety of keys (Richardson and Laroche, 1979; Laroche 
and Richardson, 1980, 1981; Matarese et ah, 1989; 
Laidig and Adams, 1991; Moser, 1996) and measured 
for standard length (SL, to the nearest mm). Densities 
for visually identified species were calculated by divid¬ 
ing catch by total distance towed (km) and assuming 
a relatively constant mouth opening for all the tows. 
Because of ambiguous and overlapping meristics, most 
(96%) late-larval and juvenile rockfishes could be iden¬ 
tified only to the genus Sebastes and were stored in 
ethanol for genetic analysis. 
DNA extraction and data collection 
To perform genetic analysis, we took samples from cau¬ 
dal fin tissue of late-larval and juvenile rockfish (2534 
individuals) collected in 144 different tows of the trawl. 
Samples were randomly selected from among the 96% 
of samples that could not be identified visually. On av¬ 
erage, 26% of the individual rockfish from each haul 
were sampled for genetic material to obtain a repre¬ 
sentative sample of catch. Total genomic DNA was ex¬ 
tracted by using a glass-fiber plate extraction protocol 
(Ivanova et al., 2006). Polymerase chain reaction (PCR) 
was used to amplify a 782-base-pair (bp) fragment of 
the mitochondrial DNA (mtDNA) cytochrome b gene by 
using previously published GluRF and CB3RF primers 
(Rocha-Olivares et al., 1999). Thermal cycling condi¬ 
tions included an initial denaturation at 94°C for 2.5 
min, followed by 35 cycles of 94°C for 45 sec, 56°C for 
1.5 min, and 72°C for 1.5 min. Final extension was car¬ 
ried out at 72°C for 3 min. PCR products were cleaned 
using a procedure with ExoSap-IT 1 reagent (Thermo 
Fisher Scientific, Waltham, MA), and were cycle se- 
1 Mention of trade names or commercial companies is for iden¬ 
tification purposes only and does not imply endorsement by 
the National Marine Fisheries Service, NOAA. 
