GSM9689405: Sample-51-6; Daphnia magna; RNA-Seq
We removed Daphnia for transcriptomic analyses from their treatments without being measured to minimize stress. We stored them individually in 500 μL of RNAlater and immediately flash froze them in liquid nitrogen. We then stored them at -80o F until RNA extraction. We extracted RNA from individual Daphnia using a RNeasy Micro Kit (Qiagen) according to the manufacturer's instructions and the extracted RNA was stored in -80o F until sequencing by RNA-seq. The rRNA depletion procedure was used in RNA-Seq library preparation for total of 84 daphnia RNA samples. Seven hundred ng of total RNA from each sample was subjected to library preparation using NEBNext Ultra II Directional RNA Library Prep with NEBNext rRNA Depletion Kit (New England Biolab, NEB, Ipswich, MA, USA) followed by the manufacturer's protocol. Each library sample (total of 84) was individually barcoded with NEBNext Dual Index Primers (NEB, Ipswich, MA, USA). The QC of each library was validated by using Qubit fluorometer with Qubit dsDNA HS Assay Kit (Life Technologies, Grand Island, NY) and the Agilent Bioanalyzer 2100 system with Agilent DNA 1000 kit (Agilent Technologies, Santa Clara, CA).