BSA11S Dark oxic transcriptome Replicate 3
RNA was extracted using the MasterPure Complete DNA and RNA Purification Kit (Lucigen, Middleton, WI). The RNA purification protocol was followed after resuspension of frozen pellets in 600 L tissue and cell lysis solution containing 2 L proteinase K. After DNA removal, RNA was resuspended in 35 L TE Buffer. A Zymo-Seq RiboFree total RNA library preparation kit (Zymo Research Corp., Irvine, CA) was used for library preparation 10 (library type: fr-firststrand, fr-secondstrand). The quantity and quality of extracted RNA was determined using an Agilent 2100 Bioanalyzer RNA assay (Agilent Technologies, Santa Clara, CA) or Caliper (PerkinElmer, Waltham, MA). Final libraries were checked using a Qubit 2.0 fluorometer (Invitrogen, Carlsbad, CA), Agilent Bioanalyzer DNA assay, or Caliper assays (PerkinElmer, Waltham, MA). Libraries were then prepared for sequencing in single-end 150 bp mode on a NextSeq 500 system (Illumina, San Diego, CA).