GSM9032669: prostate_tissue_11_BPH_biol_rep_3; Canis lupus familiaris; RNA-Seq
RNeasy Fibrous Tissue Mini Kit (Qiagen) according to the manufacturer's instructions with on column Dnase digestion The library was prepared using 350 ng of total RNA and the QuantSeq 3' mRNA library preparation FWD kit (Lexogen) according to the manufacturer's instructions. Briefly, the first strand of cDNA was synthesized, RNA was removed, and the second strand was synthesized using Unique Molecular Identifiers (UMI). Library preparation encompassed 13 cycles of PCR enrichment (initial denaturation: 98 °C, 30 sec; 13 cycles of 98 °C for 10 sec, 65 °C for 20 sec, 72 °C for 30 sec; final elongation: 72 °C, 1 min) followed by purification of the amplified product. Subsequent to this, a quality control assessment of the libraries was conducted using the Bioanalyser High Sensitivity DNA chip (Agilent Technologies). Thereafter, the libraries were quantitatively assessed by qPCR in accordance with the Illumina protocol. This facilitated the equimolar pooling of the libraries, which were subsequently sequenced on the NextSeq500 (Illumina) instrument using single reads of 75 base pairs on a high-output configuration.