Yucesan Emrah, Ozten Nur
Institute of Life Sciences and Biotechnology, Bezmialem Vakif University, Istanbul, Turkey.
Department of Pharmaceutical Toxicology, Faculty of Pharmacy, Bezmialem Vakif University, Istanbul, Turkey.
Methods Mol Biol. 2019;2054:137-145. doi: 10.1007/978-1-4939-9769-5_9.
Genome sequencing methods have basically similar algorithms, although they show a few differences between the platforms. The human genome contains approximately three billion base pairs, and this amount is huge and therefore impossible to sequence at one step. However, this amount is not a problem for developed technology. Researchers break DNA into small random pieces and then sequence and reassemble. Library preparation, sequencing, bioinformatic approaches and reporting. High-quality library preparation is critical and the most important part of the next-generation sequencing workflow. Successful sequencing directly requires high-quality libraries. Sequencing is second step and all high-throughput sequencing approaches are generally based on conventional Sanger sequencing. After preparation of library and sequencing, later steps are completely computer-based (in silico) approaches called as bioinformatics.
基因组测序方法的算法基本相似，尽管不同平台之间存在一些差异。人类基因组包含大约30亿个碱基对，这个数量非常庞大，因此不可能一步完成测序。然而，对于已开发的技术来说，这个数量并不是问题。研究人员将DNA随机切割成小片段，然后进行测序和重新组装。包括文库制备、测序、生物信息学方法和报告。高质量的文库制备至关重要，是下一代测序工作流程中最重要的部分。成功的测序直接依赖于高质量的文库。测序是第二步，所有高通量测序方法通常都基于传统的桑格测序。在文库制备和测序之后，后续步骤完全是基于计算机（电子）的方法，即生物信息学。
