design: We conducted high throughput sequencing of the bacterial 16S rRNA gene from fecal material of wild-caught slender anoles prior to testing of their heat tolerance and heat tolerance plasticity. Total DNA was extracted from feces using a Zymo Research Quick-DNA HMW MagBead Kit. A two-step PCR protocol was used to amplify the V4 region of the bacterial 16S rRNA gene and add dual-index barcodes. Sequencing was then conducted by the Naos Molecular Laboratory at the Smithsonian Tropical Research Institute in Panama City, Panama.