GSM5025020: wild type (cwo5073 control) ZT18 rep1; Drosophila melanogaster; RNA-Seq
Total RNA was isolated using Trizol (Invitrogen), treated with a TURBO™ DNase (Thermo Fisher Scientific), precipitated and purified with Lithium Chloride (Thermo Fisher Scientific) following the manufacture's instructions. 1.0 μg of total RNA was used to isolate mRNA using the NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs), which was used for RNA library construction with the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina (New England Biolabs). The resulting cDNA products were then used as template for PCR amplification for 12 cycles following the PCR conditions in the manufacturer's instructions, and after purification the eluted DNA fragments were sent for sequencing. RNA (cDNA) libraries were mixed and multiplexed at the same equimolar concentrations and sequenced using Illumina Next Seq 500 system using 75bp single end reads