title: GSM9013615: NPp53T castrated biol rep 4; Mus musculus; RNA-Seq
library_construction_protocol: Single prostates were dissected in cold 1X PBS and split into halves containing single anterior, dorsal, lateral and ventral prostate lobes. A FastPrep 24 5G bead grinder (MP Biomedicals) with stainless steel beads were used to homogenenize tissues for RNA extraction using the MagMax96 for Microarrays Total RNA Isolation kit following the Spin procedure. For castrated tissues, extracted RNA was treated with DNase. Total RNA samples were quantified and qualified using an Agilent Bioanalyzer RNA Nano kit.  Samples were prepared for sequencing using the Illumina Stranded Total RNA Prep, Ligation with Ribo-Zero Plus.  Inputs for hormonally intact samples were normalized to 100 ng; inputs for castrated samples were normalized to 500 ng.  Indexed adapters from IDT for Illumina RNA UD Indexes Set A were used for both projects.   Libraries for hormonally intact samples were sequenced using an Illumina NextSeq 500, with 2x76 bp reads; libraries for castrated samples were sequenced using an Illumina NextSeq 2000, with 2x100 bp reads, at a read depth of 10-15M paired-end reads/sample.