GSM4781205: SkMuscle_P_12mo_rep4; Rattus norvegicus; RNA-Seq
Total RNA was isolated from gastrocnemius muscle using TRIzol® reagent (Invitrogen, Carlsbad, CA) following manufacturer's protocol. For each metal and time point, three Affymetrix chips (Clariom S Rats) were used with 250 ng of total RNA derived from a pooled sample from 2-3 animals. Library preparation and sequencing were performed by Novogene Bioinformatics Technology Corporation. Briefly, RNA sequencing libraries were constructed with TruSeq Stranded Total RNA Sample Prep Kits (Illumina, San Diego, CA) using 1000 ng of total RNA. Samples were indexed with adaptors and submitted for paired-end 2 × 100-bp sequencing using an Illumina HiSeq 2000 instrument. Sequencing was performed on ten RNA samples in each lane of the flow cell, following the manufacturer's instructions.