756 



Fishery Bulletin 100(4) 



the recognition sequence (bp) and the number of recogni- 

 tion sites identified by each enzyme were the following: for 

 ND5/6— Ase I (6, 5},Ava I (6, 4), Msp I (4, 4),Rsa 1(4, 5); for 

 cytochrome b—Alu I (4, 2), Hae III (4, 2), Mse I (4, 7); and 

 for cytochrome oxidase — Alu I (4, 4). A total of 150 nucleo- 

 tides were surveyed for polymorphism or approximately 

 3.5'7f of the combined 4300-bp region. 



Microsatellites Thirteen microsatellite loci were screened 

 for polymorphism, amplification quality, and null alleles in 

 walleye pollock by using primer pairs derived from Atlan- 

 tic cod, Gadus morhua, (Gmol, Gmo2, Gmo9, GmolO, 

 Gmol23, Gmol32, and Gmol45; Brooker et al., 1994) and 



walleye pollock (Tch.5, TchlO, Tchll. Tchl2. Tchl8, and 

 Tch22; O'Reilly et al., 2000). PCRs were performed in 10 

 pL volumes containing 10 mM Tris-HCl (pH 8.3), 50 mM 

 KCl, 1.5 mM MgCU, 0.8 mM dNTPs, 0.05 units/pL Taq 

 polymerase, 0.3-1.2 pM primer, and about 250 ng DNA 

 template. The following PCR profile was used: 92°C (2 

 min) -I- 30 cycles of (92°C (30 sec) + X°C (30 sec) + 72°C 

 (140 sec)) + 72°C (5 min) where the annealing tempera- 

 ture X varied among microsatellites. 



Microsatellites were size fractionated by using an Ap- 

 plied Biosystems Inc. (ABI, Foster City, (ilA) 377-96 au- 

 tomated DNA sequencer operated in GeneScan^'^' mode 

 (ABI, 1996a). Data were analyzed by using the internal 



