NextSeq 500 sequencing; Lymph miRNA in FcgRIIA transgenic mice, in K/BxN rheumatoid arthritis
Lymph miRNA in FcgRIIA transgenic mice, in K/BxN rheumatoid arthritis
Thoracic lymph was collected as described in Milasan, Tessandier et al, JEV, 2016 (PMID: 27664155). Cells were removed by centrifugation (400 g, 5 min), and acellular lymph was stored at -80C up to RNA extraction. RNA was extracted using standard TriZol LS protocol. Total RNA of each sample was used to prepare the miRNA sequencing library which included the following steps: 1) 3'-adapter ligation with T4 RNA ligase 2 (truncated); 2) 5'-adapter ligation with T4 RNA ligase; 3) cDNA synthesis with RT primer; 4) PCR amplification; 5) extraction and purification of ~130-150 bp PCR amplified fragments (correspond to ~16-30 nt small RNAs) from the PAGE gel.
Experimental Factor: disease
Control (PBS injection)