We sought to obtain representative sampling from the brown anoles native range, and a set of areas where large, introduced populations now occur. For the native range, we obtained samples from Cuba through museum loans (MVZ and MCZ) and from the Bahamas and Cayman Islands through a collaboration with Dr. Amber Wright. For the SE US introduced range (Florida and Alabama), we obtained samples through museum loans (LACMA, FLMNH, MVZ), and obtained samples from Tampa, FL through a collaboration with Dr. Amber Wright. Samples were collected in Orange County, California through previous survey efforts (Fisher et al. 2020). We surveyed and sampled the islands of Hawaii, Maui, Molokai, Lanai, Oahu, and Kauai in the Hawaiian Archipelago. In Hawaii, specimens were collected through survey efforts and sites were selected based on personal observations of the species presence or with the help of community science records on the iNaturalist platform. From our field collections, whole specimens were formalin-fixed and livers dissected and preserved in 95% ethanol for DNA extractions. DNA was extracted from all tissues using a Qiagen DNeasy Blood and Tissue Kit (Qiagen, Inc.) and quantified using a Qubit dsDNA BR assay kit (Thermo Fisher, Waltham, MA). We prepared two genomic libraries following a modified double-digest RADseq (ddRAD) protocol of Peterson et al. (2012) (protocol in Barley et al. 2019) and sequenced on an Illumina NovaSeq SP Lane (100SR, 10X chemistry), with one library per lane. We demultiplexed each sample using the process_radtags program in STACKS V.2.514 (Catchen et al. 2013).
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