title: GSM1602986: S10; Homo sapiens; RNA-Seq
library_construction_protocol: Neat frozen (-80°C) semen samples from ejaculate obtained with permission of male partner from couples undergoing fertility treatment at two clinics - Serono(S) and Harvard(H) - were  thawed then immediately subjected to purification through a 50% gradient of Puresperm (NidaCon International AB) in order to obtain pure sperm Sperm RNA was isolated and purified using a modified RNeasy (Qiagen) protocol and finally the RNA quality assessed for DNA contamination as described in paper A total of 2 ng of unfractionated input RNA along with the recommended concentration of ERCC RNA spike-in controls (Life Technologies) was reverse transcribed and the resulting cDNA amplified using SeqPlex RNA Amplification (Sigma-Aldrich Co.) allowing for the use of samples with low quantity of spermatozoa Amplified primer dimers were removed prior to library construction using Agencourt AMPure XP (Beckman Coulter) and libraries were constructed with 50 ng of amplified cDNA using NEBNext Ultra DNA Library Prep Kit for Illumina (New England Biolabs) kit. 6bp inline Illumina index primers allowed for multiplexing of samples (Index in file name) Paired-end sequencing (2x51  + 6Index) was performed  using an Illumina HiSeq 2500 sequencer
attributes.tag: GEO Accession
attributes.value: GSM1602986