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Fishery Bulletin 107(4) 
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Figure 1 
Map of sampling locations for mutton snapper (Lutjanus analis) collected across Florida and the 
Caribbean for microsatellite analysis. Stars denote sampling sites. Collection dates were as fol- 
lows: Belize = May 2003; Dry Tortugas, FL = June-December 2003; Honduras=April 2004; Jupiter, 
FL = October 2004; and Puerto Rico = September 2004-February 2005. Adult fish were collected with 
hook and line; juvenile fish (Jupiter site only) were collected by seining. 
wrasse (Thalassonia bifasciatum) lacks structure even 
at the scale of the entire Caribbean basin. Similarly, the 
slippery dick ( Halichoeres bivittatus ) shares mtDNA hap- 
lotypes across biogeographical provinces and locations 
separated by more than 2000 km (Rocha et al., 2005). 
The results of these studies indicate similar oppor- 
tunities for evaluating patterns of connectivity among 
reef fish populations in the Dry Tortugas, southeast 
Florida, and the Caribbean basin. In July 2001, the 
Tortugas South Ecological Reserve (TSER) was estab- 
lished approximately 110 km southwest of Key West, 
Florida, encompassing an historical fishing area known 
as Riley’s Hump. This topographic feature was instru- 
mental in the delineation of the reserve boundaries 
because it serves as a spawning site for various com- 
mercially and recreationally important snapper and 
grouper species (Lindeman et al., 2000). The high site 
fidelity and temporal stability exhibited by these spawn- 
ing aggregations (Domeier and Colin, 1997) has led to 
their heavy exploitation and rapid decline on Riley’s 
Hump (Burton, 2002). In fact, this site represents the 
last known major spawning aggregation for the mutton 
snapper (Lutjanus analis) in U.S. waters, making this 
species of particular interest to both conservationists 
and fishery managers. 
In this study, we used L. analis as a focal species 
to examine eight high-resolution genetic markers to 
estimate connectivity among populations in the TSER, 
in southeast Florida, and at other sites throughout the 
Caribbean, with the ultimate goal of identifying larval 
source populations. 
Materials and methods 
Sample collection and genotyping 
Mutton snapper tissue samples were obtained between 
May 2003 and February 2005 from five geographic loca- 
tions and stored in salt-saturated dimethyl sulfoxide 
(DMSO). Samples of adult fish came from Gladden Spit, 
Belize (BZ); Roatan, Honduras (HN); Dry Tortugas (DT), 
Florida; and Mayaguez, Puerto Rico (PR) (Fig. 1); in 
addition, a sample of juvenile fish (standard length [SL]: 
34-233 mm) from Jupiter (JP), Florida, was collected 
from Jupiter Inlet in October 2004 to serve as a down- 
stream population. Genomic DNA was isolated according 
to a modification of the “rapid isolation of mammalian 
DNA” protocol of Sambrook and Russell (2000). 
Two-hundred and forty-five individuals were geno- 
typed at eight microsatellite loci (Table 1). Amplifi- 
cations (15 pL) contained 5-20 ng template DNA, 15 
mM Tris-HCl, pH 8.0, 50 mM KC1, 1.5 mM MgCl 2 , 
2.5 mM each dNTP, 0.5 uM each primer, and 0.75 U 
