GSM2232877: cMPG_4c; Macaca fascicularis; RNA-Seq
at different treament time points cells were resuspended in QIAzol for cell lysis and the lysate was processed with miRNeasy Mini kit combined with DNase treatment on a solid support for RNA extraction (Qiagen Inc., USA). RNA quality with RIN values > 9.7 was determined by electrophoresis on Agilent RNA 6000 Nano Bioanalyzer microfluidic chips. For RNA-sequencing library preparation, 500 ng total RNA was used as input. First, mRNA was enriched using Oligo-dT-containing capture beads using the Illumina TruSeq Stranded mRNA Sample Prep Kit (Illumina #RS-122-2101). Enriched mRNA was resuspended in TruSeq Fragmentation buffer and reversed transcribed into cDNA using random hexamer primer. Then cDNA was further processed for the construction of barcoded sequencing libraries according to the manufacturer's recommendations using the TruSeq Stranded mRNA Sample Prep Kit (Illimina #RS-122-2101). 6 barcoded samples were pooled to fit onto 1 lane of an Illumina flow cell containing in total 8 lanes. Sequencing was performed with the Illumina TruSeq SBS Kit v4-HS chemistry (Illumina #FC-401-4003) on an Illumina HiSeq2500 instrument with 50 cycles of 2x50 bp paired-end sequencing. barcode: CGTACG