miRNAseq_Blood

This file contains the small RNA-seq dataset that was used in the unpublished manuscript and was further analyzed with the DEGviewer online software. The dataset shows significant miRNA fold changes when comparing CTE-induced groups with the control group.RNA isolated from each sample was used to construct sequencing libraries using the SMARTer smRNA-Seq Kit for Illumina following the manufacturer's protocol. Briefly, the Input RNA was polyadenylated to provide a priming sequence for the oligo-(dT) primer. cDNA synthesis is primed by the 3’ smRNA dT Primer, which incorporates an adapter sequence at the 5’ end of each first-strand cDNA molecule. When the MMLV-derived PrimeScript™ Reverse Transcriptase (RT, cat no: 2680. Takara Bio, Otsu, Japan) reaches the 5’ end of each RNA template, it adds nontemplated nucleotides which are bound by the SMART smRNA Oligo-enhanced with locked nucleic acid (LNA) technology for greater sensitivity. In the template-switching step, PrimeScript RT uses the SMART smRNA Oligo as a template for the addition of a second adapter sequence to the 3’ end of each first-strand cDNA molecule. In the next step, full-length Illumina adapters (including index sequences for sample multiplexing) are added during PCR amplification. The Forward PCR Primer binds to the sequence added by the SMART smRNA Oligo, while the Reverse PCR Primer binds to the sequence added by the 3’ smRNA dT Primer. The resulting cDNA library included the sequences required for clustering in an Illumina flow cell. Libraries were validated by checking their size, purity, and concentration using an Agilent Bioanalyzer. The libraries were pooled in equimolar amounts and sequenced using an Illumina NovaSeq instrument. Image decomposition and quality value calculations were performed using Illumina pipeline modules by Macrogen, Inc. (Daejeon, South Korea).