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Fishery Bulletin 91|4). 1993 



analysis on these two DNA fragments within and be- 

 tween species. 



Materials and methods 



Sample collection 



The species used and sources of collection are listed in 

 Table 1. All of the fresh or frozen specimens were col- 

 lected in the Miami and Key West, FL, area during 

 1990 and 1991. Fertilized eggs and hatched larvae of 

 Lutjanus synagris were raised at 28°C in 380-L tanks 

 and fed a combination of cultured rotifers and wild- 

 caught zooplankton. The eggs (4 to 6 hours after in- 

 semination) and larvae (3 and 9 days old) were fixed 

 with 95% ethanol and kept for two months at room 

 temperature. Two eggs and two larvae were randomly 

 chosen. All of the museum specimens were obtained 

 from the Ichthyology Museum at the University of Mi- 

 ami. The time since preservation ranged from 5 to 42 

 years and it is likely that most were fixed with formal- 



dehyde first and then transferred to ethanol or iso- 

 propyl alcohol (C. R. Robins, Professor, Rosenstiel School 

 of Marine and Atmospheric Science, Division of Marine 

 Biology and Fisheries, Univ. Miami, 4600 Rickenbacker 

 Causeway, Miami, FL 33149-1098, pers. commun.). How- 

 ever, exact records on preservation methods were not 

 available. Samples are labelled as to source, for example, 

 Lcyl and LcyMl designate Lutjanus cyanopterus fresh 

 or frozen specimen No. 1 and L. cyanopterus museum 

 specimen No. 1, respectively. 



DNA extraction 



Fresh or frozen specimens Mitochondria-enriched or 

 total genomic DNA samples were prepared from fresh 

 or frozen specimens by the method of Chapman and 

 Powers ( 1984) with slight modification. 



Museum specimens A piece of white muscle (0.1- 

 0.2 g) was dissected from each individual, shredded 

 and rinsed in TEK buffer (50mM Tris, 10 mM EDTA, 



