GSM7071096: Nfr Muscle Old Het Fasted F Sample 21; Nothobranchius furzeri; RNA-Seq
Samples were disrupted by bead beating in 300μl of TriZol (Sigma) and a single 3mm metal bead (Eldan, BL6693003000) using TissueLyzer LT (QIAGEN, #85600) with a dedicated adaptor (QIAGEN, #69980). Beating was performed twice at 50Hz for 2 minutes. RNA extraction was performed with Direct-zol RNA Purification Kits (Zymo). RNA concentration and quality were determined by using an Agilent 2100 bioanalyser (Agilent Technologies). Library preparation was performed using KAPA Stranded mRNA-Seq Kit (ROCHE-07962193001) according to the recommended protocols. Library quantity and pooling were measured by Qubit (dsDNA HS, Q32854), Size selection at 4% agarose gel. Library quality was measured by Tape Station (HS, 5067-5584). Libraries were sequenced by NextSeq 2000 P3, 50 cycle, 70 bp single-end (Illumina, 20046810) with ~35 million reads per sample.