Determining the sequence of DNA and RNA molecules has a huge impact on the understanding of cell biology and function. Recent advancements in next-generation short-read sequencing (NGS) technologies, drops in cost and a resolution down to the single-cell level shaped our current view on genome structure and function. Third-generation sequencing (TGS) methods further complete the knowledge about these processes based on long reads and the ability to analyze DNA or RNA at single molecule level. Long-read sequencing provides additional possibilities to study genome architecture and the composition of highly complex regions and to determine epigenetic modifications of nucleotide bases at a genome-wide level. We discuss the principles and advancements of long-read sequencing and its applications in genome biology.

确定DNA和RNA分子的序列对细胞生物学和功能的理解有巨大影响。下一代短读测序（NGS）技术的最新进展，成本下降以及分辨率降至单细胞水平，这形成了我们当前对基因组结构和功能的看法。第三代测序（TGS）方法基于长期阅读以及在单分子水平上分析DNA或RNA的能力，进一步完善了有关这些过程的知识。长时间阅读测序为研究基因组结构和高度复杂区域的组成以及在全基因组水平确定核苷酸碱基的表观遗传修饰提供了其他可能性。我们讨论了长读测序的原理和进展及其在基因组生物学中的应用。