title: GSM8186880: sample90, heart, ZSF1 Obese, BT2, 3 Study A, rep10; Rattus norvegicus; RNA-Seq
library_name: GSM8186880
library_construction_protocol: Extraction was performed using the RNeasy Plus Universal Mini Kit (Qiagen) on approximately 50mg of frozen tissue. Total RNA was evaluated for quality (RIN scores ≥ 7) by 4200 TapeStation System (Agilent) using RNA ScreenTape (cat. 5067-5576, Agilent). The concentrations were measured by High Lunatic plate (cat. 7012000, Unchained Labs). 400 ng total RNA was used as input for the mRNA-Seq library preparation using the TruSeq Stranded mRNA Library Prep Kit (cat. 20020594, Illumina). Poly-A containing mRNA was purified using poly-dT beads, fragmentation of RNA, first strand synthesis, second strand cDNA synthesis and End repairing, A-tailing, Adapter ligation, followed by PCR amplification with 15 PCR cycles to generate final cDNA libraries. The final libraries were then evaluated for quality by 4200 TapeStation System (Agilent) using D1000 ScreenTape (cat. 5067-5582, Agilent). The concentrations were measured using Qubit 1X dsDNA HS Assay Kit (cat. Q33231, Invitrogen-Life Technologies Corporation). The libraries were then normalized to 4 nM and re-quantitated using Qubit 1X dsDNA HS Assay Kit (cat. Q33231, Invitrogen- Life Technologies Corporation) and then pooled into two pools for sequencing. The pools were loaded on two NextSeq2000 sequencers for 75 Paired End run using P3 200 cycle kit (cat. 20040560, Illumina).