<b>Small RNA Sequencing</b><b> (Mouse Blood)</b>

This file contains the small RNA-seq dataset that was used in the unpublished manuscript. 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).

本文件包含用于未发表手稿的小RNA测序（small RNA-seq）数据集。该数据集显示，与对照组相比，慢性创伤性脑病（CTE）诱导组的微小核糖核酸（miRNA）表达倍数变化具有显著性差异。从每个样本中分离的RNA按照制造商的方案，使用Illumina SMARTer小RNA测序试剂盒构建测序文库。简要步骤如下：输入RNA经多聚腺苷酸化处理，为寡聚脱氧胸苷（oligo-(dT)）引物提供结合序列。互补脱氧核糖核酸（cDNA）合成由3’小RNA dT引物引发，该引物在每条第一链cDNA分子的5’端引入接头序列。当源自莫洛尼鼠白血病病毒（MMLV）的PrimeScript™逆转录酶（RT，货号：2680，Takara Bio，日本大津）到达每个RNA模板的5’端时，会添加非模板核苷酸，这些核苷酸与经锁核酸（LNA）技术增强的SMART小RNA寡核苷酸结合，以提高灵敏度。在模板转换步骤中，PrimeScript逆转录酶以SMART小RNA寡核苷酸为模板，在每条第一链cDNA分子的3’端添加第二个接头序列。下一步，在聚合酶链式反应（PCR）扩增过程中添加全长Illumina接头（包括用于样本多重化的索引序列）。正向PCR引物结合SMART小RNA寡核苷酸引入的序列，而反向PCR引物结合3’小RNA dT引物引入的序列。所得cDNA文库包含在Illumina流动槽中进行聚类所需的序列。通过Agilent Bioanalyzer检测文库的大小、纯度和浓度以进行验证。文库按等摩尔量混合，使用Illumina NovaSeq仪器测序。图像分解和质量值计算由Macrogen公司（韩国大田）使用Illumina pipeline模块完成。