GSM9140967: OPE 2um #1; Danio rerio; RNA-Seq
TRIzol reagent protocol The libraries were prepared for 150bp paired-end sequencing using TruSeq Stranded mRNA Sample Prep Kit. Namely, mRNA molecules were purified and fragmented from 0.1-1μg of total RNA using oligo (dT) magnetic beads. The fragmented mRNAs were synthesized as single-stranded cDNAs through random hexamer priming. By applying this as a template for second strand synthesis, double-stranded cDNA was prepared. After sequential process of end repair, A-tailing and adapter ligation, cDNA libraries were amplified with PCR (Polymerase Chain Reaction). Quality of these cDNA libraries was evaluated with the Agilent 2100 BioAnalyzer (Agilent, CA, USA). They were quantified with the KAPA library quantification kit (Kapa Biosystems, MA, USA) according to the manufacturer's library quantification protocol. Following cluster amplification of denatured templates, sequencing was progressed as paired-end (2×150bp) using Illumina Novaseq 6000 (Illumina, CA, USA).