GSM4649301: Bovine endometrium_2_sPIF; Bos taurus; RNA-Seq
Total RNA was extracted using the Norgen Total RNA purification plus kit following the manufacturer's instructions, with the addition of the use of a bead beater with the lysis buffer to aid tissue disruption. Total RNA was prepared for sequencing using the TruSeq v2 RNA-Seq kit, using the manufacturer's protocol, up to the validate library step. Following the enrichment of cDNA fragments with adapters, the cDNA was quantified using a Qubit 2.0, dsDNA broad range assay. Each sample was diluted to 10 nM with 10 nM Tris HCl and 0.5 % Tween-20 in nuclease free water. Samples were pooled and subsequently diluted to 2 nM with elution buffer (Qiagen, Manchester, UK) and then to 1 nM with 0.1 M sodium hydroxide, before being held at room temperature for 5 minutes to denature the DNA. Following denaturation, the samples were diluted to 10 pM in hybridisation buffer and loaded onto a cBot to cluster cDNA onto two lanes of a V4 High output flow cell. Samples were paired-end sequenced on an Illumina HiSeq 2500 and base pairs per read were set to 126 bp. Samples 9-14 were sequenced twice due to low reads due to the sequencing process compared to samples 1-8
GEO Accession
GSM4649301