title: GSM3061458: Pregnant glandular epithelium mare M; Equus caballus; RNA-Seq
library_construction_protocol: Laser capture microdissection (LCM)  Stained frozen sections (10 µm) were prepared using a microtome cryostat CryoStar NX50 (Histocom AG, Zug, Switzerland) on PEN-Membrane Glass Slides®, and Arcturus® HistoGene® Frozen Section Staining Kit (Applied Biosystems). The collection of luminal epithelium (LE), glandular epithelium (GE) and stromal areas (S) was performed using an ArcturusXT® Laser Capture Microdissection instrument.  RNA isolation, quantification and quality control   RNA isolation was performed with the Arcturus PicoPureTM RNA Isolation Kit (Thermo Fisher Scientific, Darmstadt, Germany). Concentration of the RNA was measured with a QuantusTM Fluorometer (Promega; sensitivity 390 pg RNA/ml using QuantiFluor® RNA Dye). Quality was analyzed by the use of the Agilent 2100 Bioanalyzer (Eukaryote Total RNA Pico Assay), which estimates the concentration of the RNA sample and provides a quality score (RNA integrity number, RIN). For the generation of RNA-Seq libraries from LCM samples, the Ovation® Single Cell RNA-Seq System (NuGEN Technologies, Inc., European office, Leek, The Netherlands) was used starting from 1 ng total RNA. Since for a number of libraries the concentration of the obtained cDNA fragments was too low (Agilent Bioanalyzer DNA High Sensitivity assay), an additional amplification was performed with the KAPA HiFi Library Amplification Kit (Axon Lab AG, Baden, Switzerland). Single-end reads (125 bp) were generated on an Illumina HiSeq 2500 instrument running multiplexed libraries on two lanes of one flow cell.
attributes.tag: GEO Accession
attributes.value: GSM3061458