alias: GSM3532111
title: GSM3532111: Blood_PBS_rep2; Mus musculus; RNA-Seq
library_construction_protocol: Blood was collected post mortem via heart puncture on day two after infection (n = 3 mice/treatment) and kept at -20 °C in RNAlater (Thermo Fisher Scientific). RNA was extracted using the Mouse RiboPure Blood RNA Isolation Kit and globin RNA depleted using the GLOBINclear Kit, mouse/rat (Thermo Fisher Scientific). Quality and amount of RNA were determined using the Qubit RNA BR Assay Kit with Qubit 2.0 (Thermo Fisher Scientific) and Agilent RNA 6000 Nano Kit on the Agilent 2100 Bioanalyzer. Only samples with RIN score above 7 were further analyzed. Libraries were prepared using NEBNext Ultra Directional RNA Library Prep Kit for Illumina, NEBNext® Multiplex Oligos for Illumina® (Index Primers Set 1 and 2) and NEBNext Poly(A) mRNA Magnetic Isolation Module (New England Biolabs). Library quantification was performed by real-time PCR using the KAPA Library Quantification Kit - Illumina Platforms - Complete kit (Universal) and KAPA Library Amplification Kit - Real-time PCR library amplification, with fluorescent standards (KAPA Biosystems). Libraries were sequenced twice on the Illumina Nextseq platform (Illumina) using V1 reagents to a total read depth of 60 million paired-end, 75 base-pair reads.
identifiers.namespace: GEO
identifiers.id: GSM3532111