Development and characterization of 11 microsatellite loci for the Mona Island iguana (Cyclura cornuta stejnegeri)

Abstract

We isolated and characterized 11 microsatellite loci in the Mona Island iguana (Cyclura cornuta stejnegeri). Eleven loci exhibit moderate to high allelic diversity (two to 12 alleles, mean = 4.5) and polymorphism (mean observed heterozygosity, 0.56; range, 0.26 to 0.78) in 41 adults. This marker set has low probability of identity and high parentage exclusion power and will be suitable for studies of paternity, social organization and relatedness in this species.

Age‐dependent, negative heterozygosity–fitness correlations and local effects in an endangered Caribbean reptile, Iguana delicatissima

“…To identify polymorphic microsatellite loci in I. delicatissima , 60 anonymous microsatellites identified in studies of Cyclura species (An et al., 2004; Lau et al., 2009; Rosas et al., 2008; Welch et al., 2011) were tested for amplification using a basic PCR touchdown cycle (Don, Cox, Wainwright, Baker, & Mattick, 1991). Sequence amplification was verified on 1% agarose gels.…”

“…DNA was extracted from blood samples using either an ABI PRISMᵀᴹ 6100 Nucleic Acid Prep Station (Applied Biosystems, Foster City, California, USA) or a Maxwell® 16 Nucleic Acid Extraction System with a Maxwell® 16 Tissue DNA Purification Kit (Promegaᵀᴹ). To identify polymorphic microsatellite loci in I. delicatissima, 60 anonymous microsatellites identified in studies of Cyclura species (An et al, 2004;Lau et al, 2009;Rosas et al, 2008;Welch et al, 2011) were tested for amplification using a basic PCR touchdown cycle (Don, Cox, Wainwright, Baker, & Mattick, 1991). Sequence amplification was verified on 1% agarose gels.…”

Twenty-nine polymorphic microsatellite loci in Cyclura carinata, the Turks and Caicos Iguana, a critically endangered island endemic

“…Available molecular tools are inadequate for their identification. We characterize microsatellite loci developed for C. carinata, and identify microsatellite primers developed for Cyclura nubila , C. pinguis (Lau et al 2009), and C. cornuta (Rosas et al 2008) that cross-amplify, and are variable in C. carinata.…”

Genetic structure at three spatial scales is consistent with limited philopatry in Ricord's Rock Iguanas (Cyclura ricordii)

“…Samples were genotyped at 14 microsatellite loci found to be variable for C. ricordii . The polymorphic markers used were as follows: Ccste05 (Rosas et al, ); CIDK135, CIDK144, and CIDK184 (Welch et al, ); Z106, Z148, Z154, and Z494 (Junghwa et al, ); and D1, D11 D101, D110, D111, and D140 (Lau et al, ). CIDK135 locus (Welch et al, ) was eliminated from further analysis due to high null allele frequency p = 0.48 (Dakin & Avise, ; Table ).…”

“…Noncoding microsatellites were used to infer patterns of population structure and rates of gene flow. Over seventy microsatellite markers were screened and characterized for this species with the 2012 samples of hatchlings and adults (all microsatellite markers designed by Rosas et al, , Welch et al, , Junghwa et al, , and Lau et al, were included in the screening). Amplifications were conducted with a 2,720 Thermal Cycler (AB Applied Biosystems) following standardized 3‐primer PCR amplification according to Schuelke () and modified by Welch et al () in a total volume of 10 µl (i.e., 7.3 µl of ddH 2 O, 1.2 µl of master mix (10 µM of each dNTPs, 10x Tricine Taq Buffer and ddH 2 O), 0.04 µl forward primer, 0.2µl reverse primer, 0.2 µl fluorescent tagged primer, 0.4 U Taq polymerase, and 1.0 µl of DNA template).…”