GSM3755965: Biopsy 466; [Haemophilus] ducreyi; Homo sapiens; RNA-Seq
Biopsy samples immediately after collection and the extra inocula immediately after inoculation were placed in 2 mL RNAlater, incubated for 30 minutes at room temperature, and stored in RNAlater for one day at -80°C. Using a mini-Beadbeater (Biospec Products), we next homogenized the samples and extracted total RNA using a RNeasy Fibrous Tissue Mini kit according to manufacturer's instructions along with addition of lysozyme (20 ug/mL) added at the proteinase K step. The RNA was treated twice with TURBO DNA-free DNase (Ambion) following manufacturer's instructions. RNA concentrations and integrities were measured using a NanoPhotometer P330 (Implen). RNA samples were stored at -80°C until all samples were ready for sequencing. We removed 23S, 16S, and 5S rRNA before RNA-seq using the Ribo-Zero Gold rRNA Removal Kit (Epidemiology) (Epicentre Biotechnologies) following manufacturer's instructions and confirmed their removal using the Agilent 2100 Bioanalyzer. Twelve libraries (four mid-log phase growth H. ducreyi cultures, four infected sites, and four wounded sites) were constructed using the TruSeq Stranded Total RNA Library kit (Illumina) following manufacturer's instructions.
GEO Accession
GSM3755965