We tested the utility of a protocol using genetic markers that previously proved successful to identify the sex of Vespertilionid bats on tissues collected from live bats and carcasses of varying age from the Hawaiian hoary bat (Lasiurus cinereus semotus). This molecular method is based on genes unique to X and Y chromosomes in mammals and previously was used successfully on North American hoary bats (L. c. cinereus). Using genotyping, we determined the sex of 102 of 109 Hawaiian hoary bat tissue samples. This dataset includes all 73 samples from the original data release, plus 36 additional samples. NOTE: While previous versions are available from the author, all the records in previous versions can be found in version 4.0. First posted - October 17, 2018 (available from author) Revised - November 20, 2018 (version 2.0 - available from author) Revised - November 26, 2019 (version 3.0 - available from author) Revised - April 13, 2020 (version 4.0 - available from author) Revised - September 28, 2020 (version 5.0 - available from author) Revised - January 28, 2021 (version 6.0)