Next Generation Sequencing Facilitates Quantitative Analysis of colorectal cancer cell lines including SW480 and SW620

Purpose: The goals of this study are to compare the differentially expressed miRNAs between SW480 and SW620. Methods:Total RNA of two samples was used to prepare the miRNA sequencing library.The libraries were denatured as single-stranded DNA molecules, captured on Illumina flow cells, amplified in situ as clusters and finally sequenced for 50 cycles on Illumina NextSeq. Results:We found 72 miRNAs were downregulated on the basis of fold-change less than 0.5. However,total 160 miRNAs were upregulated on the basis of fold-change greater than 2. Conclusions: Our study represents the detailed analysis of SW480 and SW620, generated by RNA-seq technology. Our results show that NGS offers a comprehensive and more accurate quantitative and qualitative evaluation of miRNA content. We conclude that RNA-seq based transcriptome characterization would expedite genetic network analyses and permit the dissection of complex biologic functions. Overall design: SW480 and SW620 were generated by deep sequencing, using Illumina NextSeq.

研究目的：本研究旨在比较SW480与SW620细胞系间差异表达的微小RNA（miRNA）。
实验方法：提取两份样本的总RNA以构建miRNA测序文库。将文库变性为单链DNA分子，锚定在Illumina流动槽（Illumina flow cell）上，原位扩增为簇集，最终在Illumina NextSeq测序平台上开展50轮测序反应。
实验结果：基于折叠变化小于0.5的筛选阈值，我们发现72种miRNA表达下调；而基于折叠变化大于2的筛选阈值，共计160种miRNA表达上调。
研究结论：本研究完成了对SW480与SW620的详细转录组分析，该分析依托RNA测序技术完成。结果表明，下一代测序（Next-Generation Sequencing，NGS）可对miRNA的表达量实现全面且更为精准的定量与定性评估。我们认为，基于RNA测序的转录组表征能够加速遗传网络分析进程，并助力解析复杂的生物学功能。
整体实验设计：采用Illumina NextSeq测序平台对SW480与SW620进行深度测序。