Chen Zuoquan, Xie Lei, Tang Xi, Zhang Zhiyan
State Key Laboratory for Pig Genetic Improvement and Production Technology, Jiangxi Agricultural University, Nanchang 330045, China.
MethodsX. 2022 Dec 16;10:101969. doi: 10.1016/j.mex.2022.101969. eCollection 2023.
Although meiotic recombination is a key step shared by eukaryotes, the rate of recombination varies at different taxonomic levels. The construction of high-resolution genome-wide recombination maps will help us understand the variability patterns of recombination rates and their molecular basis. ONT sequencing technology has the characteristics of long read length, high throughput, and reasonable cost, and can be used as a data source for the construction of whole-gene recombination landscapes. In order to construct the genome-wide recombination map of an individual conveniently and accurately, we developed a method to construct the recombination landscape based on the third-generation sequencing technology, Oxford Nanopore Sequencing. Here we detail a step-by-step approach to efficiently and accurately construct genome-wide recombination maps using ONT pooled sequencing data. The main contents include compression homopolymers and alignment; acquisition of high-quality variants; estimation of recombinant molecules by the sliding window method; and construction of recombinant maps. The results of simulation data validation show that our method has high sensitivity and specificity at moderate heterozygous variant density and sequencing depth. This method provides a new way of constructing high-resolution individual genome recombination maps using long read sequences, and has important reference significance for the study of recombination rate variation.
虽然减数分裂重组是真核生物共有的关键步骤,但重组率在不同分类水平上有所不同。构建高分辨率全基因组重组图谱将有助于我们了解重组率的变异模式及其分子基础。纳米孔测序技术具有读长长、通量高、成本合理的特点,可作为构建全基因组重组图谱的数据源。为了方便、准确地构建个体的全基因组重组图谱,我们开发了一种基于第三代测序技术——牛津纳米孔测序的重组图谱构建方法。在此,我们详细介绍一种利用纳米孔混合测序数据高效、准确地构建全基因组重组图谱的逐步方法。主要内容包括压缩同聚物和比对;获取高质量变异;通过滑动窗口法估计重组分子;以及构建重组图谱。模拟数据验证结果表明,我们的方法在中等杂合变异密度和测序深度下具有较高的灵敏度和特异性。该方法为利用长读长序列构建高分辨率个体基因组重组图谱提供了一种新途径,对重组率变异的研究具有重要参考意义。