alias: GSM2464546
title: GSM2464546: 96 hpi, replicate 3; Acanthamoeba castellanii; Candidatus Protochlamydia amoebophila; RNA-Seq
library_construction_protocol: After collection of cells pellets were immediately resuspended in TRIzol reagent, followed by bead-beating (30s, 4.5 m/s, lysing matrix A tubes, MP Biomedicals). Subsequent RNA extraction was performed according to the TRIzol guidelines. Residual DNA was removed using the Turbo DNA-free Kit (Thermo Fisher Scientific). Ribosomal RNA was removed using the RiboZero Magnetic kit (Illumina) for Gram-positive bacteria as recommended by the manufacturer. To enrich for mRNA from symbiont-free amoebae, the RNA was additionally treated with Dynabeads mRNA Purification Kit (Thermo Fisher Scientific). For strand-specific cDNA library preparation, the NEBNext Ultra Directional RNA Library Prep Kit for Illumina in combination with the NEBNext Multiplex Oligos (New England Biolabs) was used.
identifiers.namespace: GEO
identifiers.id: GSM2464546