RNAseq of young killifish WT kidneys
Libraries were prepared from 500 ng total RNA. ERCC RNA Spike-In Mix 1 (Thermo Fischer) was added to the samples before library preparation. Enzymatic depletion of ribosomal RNA with the Ribo-Zero Gold rRNA Depletion kit (Illumina) was followed by library preparation with the TruSeq Stranded Total RNA sample preparation kit (Illumina). The depleted RNA was fragmented and reverse transcribed with random hexamer primers, second strand synthesis with dUTPs was followed by A-tailing, adapter ligation and library amplification (15 cycles). Next, library validation and quantification (Agilent Tape Station) were performed, followed by pooling of equimolar amounts of libraries. The library pools were then quantified using the Peqlab KAPA Library Quantification Kit and the Applied Biosystems 7900HT Sequence Detection System and sequenced on an Illumina NovaSeq6000 sequencing instrument with a PE100 protocol aiming for 50 million clusters per sample.
SUB14677547
Young WT kidney 5