GSM3466652: RNA-Seq ClkKO ZT22 rep1; Danaus plexippus; RNA-Seq
Brains free of eye photoreceptors were dissected in 0.5X RNA later (Ambion) to prevent RNA degradation, and stored at -80ºC until use. Total RNA was extracted using an RNeasy Mini kit (Qiagen). PolyA+ RNA was isolated from 2 μg of total RNA with NEBNext Oligo d(T) magnetic beads (New England Biolabs), and multiplexed libraries were prepared using the NEBNext® Ultra™ Directional RNA Library Prep Kit for Illumina and NEBNext Multiplex Oligos (New England Biolabs) and amplified with 12 PCR cycles, following the manufacturer's recommendations. Libraries quality and size distribution was verified on a Bioanalyzer, libraries were quantified by real-time quantitative PCR, and 16 multiplexed libraries were mixed in equimolar ratios and sequenced on a Hi-seq 2500 (Illumina) using 50bp single end reads.
GEO Accession
GSM3466652