title: GSM3575437: L16_1085_Rat_14_Testis; Rattus norvegicus; RNA-Seq
library_construction_protocol: Tissues were isolated from freshly sacrificed animals (< 1 hour from time of death to tissue fixation), washed in cold PBS, and stored in RNALater (Ambion) per the manufacturer's instructions. Each sample was then homogenized in a bead-mill in Trizol and total RNA was isolated by and treated with DNAse on a Qiagen miRNEasy column Libraries were prepared using the TruSeq mRNA library kit (Illumina) according to manufacturer's instructions with the following modifications: After prep, libraries were size-selected using 2% agarose gels on the PippinHT system (Sage Science) with a capture window of 300-600 bases. The size-selected material was then subjected to one additional cycle of PCR with fresh reagents and P5/P7 primers (Illumina) to ensure all library fragments were fully double-stranded. This step significantly reduced the percentage of smaller fragments co-purifying in the final gel size-selection. After PCR, libraries were size-selected a second time on the PippinHT with the same settings