title: GSM7158820: RNAseq_LN_TEM_rep1; Mus musculus; RNA-Seq
library_construction_protocol: For S. aureus RNA-seq: Ear pinna skin was split into the dorsal and the ventral sheets and placed in RPMI 1640 media supplemented with 2 mM L-glutamine, 1 mM sodium pyruvate, 1 mM non-essential amino acids, 50 μM β-mercaptoethanol, 20 mM HEPES, 100 U/ml of penicillin, 100 mg/ml of streptomycin, 0.5 mg/ml of DNAse I (Sigma-Aldrich) and 0.25 mg/ml of Liberase TL purified enzyme blend (Roche) for 90 minutes at 37C and 5% CO2. Digested ear skin sheets were homogenized using the Medicon/Medimachine tissue homogenizer system. T cells were FACsorted (BD FACS Aria, Becton Dickinson) from Foxp3-GFP reporter mice. The RNA from the sorted T cells was extracted using the RNeasy Plus Micro Kit (Qiagen) as per manufacturer instructions. Libraries were prepared using the Clontech SMARTer Ultra low input mRNA-Seq sequence kit and samples were sequenced paired end on one lane of NextSeq 550.  For DRG RNA-seq: Supernatant was removed from cells, cells were lysed with RLT Plus buffer (Qiagen) and the RNA was extracted to perform bulk RNA sequencing. Libraries were prepared using the Clontech SMARTer Ultra low input mRNA-Seq sequence kit and samples were sequenced paired end on one lane of NextSeq 550.  Libraries were prepared using the Clontech SMARTer Ultra low input mRNA-Seq sequence kit and samples were sequenced paired end on one lane of NextSeq 550.