GSM7098522: ID16017_muscle_2yr; Papio anubis; RNA-Seq
Approximately 10 mg of each frozen tissue sample was homogenized in 1ml QIAzol Lysis Reagent (79306, QIAGEN) using BeadBug 6 Microtube Homogenizer (D1036, Benchmark Scientific) with 1.5mm Zirconium beads (D1032-15, Benchmark Scientific). RNA was immediately extracted from the homogenate using QIAGEN RNeasy Universal Kit (73404, QIAGEN). RNA concentration and quality was measured using Agilent RNA 6000 Nano Kit and RNA 6000 Pico Kit (5067-1511 & 5067-1513, Agilent) on Agilent 2100 Bioanalyzer. TruSeq RNA Library Preparation Kit v2 (RS-122-2001, RS-122-2002, Illumina); TruSeq stranded mRNA Library Preparation Kit (20020595, Illumina) 96 libraries from 16 animals in batch 1 were performed using TruSeq RNA Library Preparation Kit v2 (RS-122-2001, RS-122-2002, Illumina) and were sequenced 50 base pairs, single-end using the Illumina HiSeq4000 according to manufacturer instructions with the goal of achieving at least 15 million reads per sample. Each sequencing lane contained 24 multiplexed samples from different RNA extraction batches. 498 libraries from 83 animals in batch 2 were performed using TruSeq stranded mRNA Library Preparation Kit (20020595, Illumina) and were sequenced 50 base pairs, paired-end using the Illumina NovaSeq6000 according to manufacturer instructions with the goal of achieving at least 30 million reads per sample. Each sequencing lane contained 84 multiplexed samples from different RNA extraction batches. Samples from the same animal were always in the same RNA extraction, library preparation, and sequencing batches. Samples from different animals were randomized in every processing step to avoid potential batch effects.