title: GSM8771202: Alveolar macrophage, wildtype, rep2; Mus musculus; RNA-Seq
library_construction_protocol: Total RNA was extracted from FACS-sorted mature AMs of Sdha-mKO and wildtype mice using TRIzol reagent (TR 188, MRC Inc, Cincinnati, USA). RNA quality was assessed by Agilent 2100 Bioanalyzer using the RNA 6000 Nano Chip (Agilent Technologies, Amstelveen, The Netherlands), and RNA quantification was performed using ND-2000 Spectrophotometer (Thermo Fisher Scientific Inc., DE, USA). RNA sequencing libraries were constructed using the QuantSeq 3' mRNA-Seq Library Prep Kit (Lexogen GmbH, Wien, Austria) according to the manufacturer's protocol. In brief, each 500 ng total RNA were prepared and an oligo-dT primer containing an Illumina-compatible sequence at its 5' end was hybridized to the RNA and reverse transcription was performed. After degradation of the RNA template, second strand synthesis was initiated by a random primer containing an Illumina-compatible linker sequence at its 5' end. The double-stranded library was purified by using magnetic beads to remove all reaction components. The library was amplified to add the complete adapter sequences required for cluster generation. The finished library is purified from PCR components.
identifiers.namespace: GEO
identifiers.id: GSM8771202_r1