GSM5396122: MZdnmt3aa_2 dpf_RNAseq; Danio rerio; ssRNA-seq
Tissue lysates were prepared from 30 pooled 2 dpf WT and MZdnmt3aa-/- mutant larvae by homogenization using TissueLyser LT (QIAGEN) with zirconia beads for 10 s at 50 Hz. RNA was extracted from the lysate using the PureLink® RNA Mini Kit (Thermo Fisher Scientific). RNA quality of all samples was RNA Integrity Number (RIN) > 9.0, using the 4150 TapeStation System (Agilent). Total RNA was treated using the NEBNext® Poly(A) mRNA Magnetic Isolation Module (New England Biolabs) for mRNA isolation. Strand-specific RNA-seq libraries were constructed using the NEBNext® UltraTM Directional RNA Library Prep Kit for Illumina® (New England Biolabs). The libraries with adapters were sequenced on the Novaseq 6000 (Illumina) with 2 × 150 bp paired-end reads. RNA extraction, mRNA isolation, library preparation, and sequencing were performed by Rhelixa Inc. (Tokyo, Japan).