BAC克隆高通量测序及短序列拼接方法的研究

In this research, we choose grass carp (Ctenopharyngodon idella) genomic BAC library as material,use three-dimensional mixing pool strategy for high efficiency and high-throughput sequencing of BAC clones.By using computer algorithms, we parse each BAC short sequence dataset from the mixing pool. Based on graph theory in computer science, we establish an precise assembly method for short sequence data set of single BAC clone. With this method, we do sequencing and assembling for specific clones in grass carp BAC library,so as to fill the gap areas of the grass carp genome frame diagram. This research will provide a simple new idea for genome sequencing and assembling, and have widely application prospects.

本研究以草鱼基因组BAC文库为材料，采用三维混合池策略，进行BAC克隆的高效率、高通量测序；通过计算机算法实现，从混合池中解析各个BAC克隆的短序列数据集。基于计算机图论理论，建立单个BAC克隆短序列数据集的精确拼接方法。运用这一方法，对草鱼BAC文库中的特定克隆进行测序和拼接，填补草鱼基因组框架图中的断裂区域。本研究将提供一种化繁为简的基因组测序及组装新思路，具有广泛的应用前景。

本研究以草鱼基因组BAC文库为材料，对三维混合池策略进行了深入研究，获得了BAC克隆三维混合池高通量测序技术的关键性参数；完成了草鱼带有雄性个体特异性标记的62M08BAC克隆测序；基于计算机图论理论，建立了单个BAC克隆短序列数据集的精确拼接方法，在对草鱼基因组进行了PacBio测序（三代测序技术）的基础上，结合三代测序数据实现了特定BAC克隆的精确组装。