GSM6356083: Muscle, Post-training, 2002; Homo sapiens; RNA-Seq
GSM6356083
RNA was extracted using 1.5mm zirconium beads (MidSci) and Zymo Quick-DNA/RNA microprep plus kit (Zymo Research) with on-column DNase treatment. RNA quality was checked via capillary electrophoresis on a fragment analyzer using the RNA Analysis DNF-471 kit (Advanced Analytical). The NEBNext Ultra II Directional RNA Library Prep Kit (Illumina) with NEBNext rRNA Depletion Kit (New England Biolabs) was used to prepare RNA-seq libraries, with a total of at least 500 ng input RNA from each sample. The manufacturer's protocols were followed, with the following exceptions: 40× adaptor dilutions were used, all bead incubations were done at room temp, a 4× lower concentration of index primers was used, and 10 cycles of library amplification were performed. The resulting libraries were pooled in equimolar amounts, quantified using the KAPA Library Quant Kit for Illumina (Kapa Biosystems), quality checked via capillary electrophoresis on the fragment analyzer using the NGS Analysis DNF-474 kit (Advanced Analytical), and were sequenced using paired-end, 75 base pair sequencing on the Illumina NextSeq 500 massively parallel sequencer at the University at Albany Center for functional genomics.