title: GSM5252248: d84_worker_rep2; Harpegnathos saltator; RNA-Seq
library_construction_protocol: Brains from transitioning Harpegnathos ants were dissected out in phosphate-buffered saline, placed in TRIzol (Invitrogen #15596026) and stored at -70 C until RNA extraction. Each brain was processed separately. To extract RNA, thawed brains were homogenized in TRIzol by pestle, and frozen and thawed again. Chloroform was added, followed by vigorous vortexing and centrifugation at 21,000 g for 10–15 minutes at 4 C. The aqueous phase was purified using RNA Clean and Concentrator kit (Zymo Research #R1013) following manufacturer's instructions. Extracted RNA was quantified using NanoDrop 2000 (Thermo Scientific) and RNA integrity was checked using High Sensitivity RNA ScreenTape (Agilent #5067-5579, 5067-5580, 5067-5581) on a TapeStation 2200 or 4200 (Aglient). 250–500 ng extracted RNA was used to prepare libraries with NEBNext Ultra II Directional RNA Library Prep Kit for Illumina (New England Biolabs #E7760S) following manufacturer's instructions. Libraries were quantified using Qubit dsDNA High Sensitivity Assay Kit (Invitrogen #Q32854) on a Qubit 2.0 fluorometer (Invitrogen) and fragment size distribution was checked using High Sensitivity D1000 ScreenTape (Agilent #5067-5584, 5067-5585, 5067-5587) on a TapeStation 2200 or 4200 (Aglient). Libraries were combined in two pools and sequenced in two runs of a NextSeq 500 machine (Illumina) in HighOutput mode with a 2x 150 bp configuration.
attributes.tag: GEO Accession
attributes.value: GSM5252248