GSM3954928
GSM3954928: LPP4_3; Bos taurus; RNA-Seq
RNA was extracted from the endometrial tissue samples by using Trizol (Invitrogen, Carlsbad, USA) following the manufacturer's direction, resuspended and stored at -80°C until use.  For the RNA sequencing procedure, an aliquot of RNA per sample (2 μg of total RNA) was DNase treated by using the Turbo RNase-free DNase kit (Ambion, Foster City, USA). The RNA concentration was determined with a Nanodrop spectrophotometer (ND-1000, ThermoFisher, Wilmington, USA) and the integrity of RNA (RIN number) was measured by using an Agilent Bioanalyzer 2100 model. For RNA library construction, one μg of purified RNA was re-suspended in 50 μl of water. Next, mRNA was purified through hybridization with oligo-dT and chemically fragmented. Double strand cDNA was synthesized, followed by end-repair, adaptor ligation and enrichment (amplification) of the libraries according to the TruSeq Stranded mRNA protocol (Illumina Technologies, CA, USA). Quality of each cDNA library was assessed on a 2100 Bioanalyzer (Agilent Technologies) by using the DNA 1000 Kit. The average size per double strand DNA fragments was 260 bp. Libraries were quantified by qPCR (Light Cycler 480 Roche) by using the Qiagen Library Quantification Kit, according to the manufacturer´s protocol, and normalized to a concentration of 3 nM. One equimolar pool containing the 12 samples was prepared from the libraries to generate the clusters in the sequencing cell. Sequencing was performed on a Hiseq 1500 system (Illumina Inc. CA, USA) generating paired-end 2x150bp reads.
GEO
GSM3954928
GEO Accession
GSM3954928