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De novo bacterial genome sequencing: Millions of very short reads assembled on a desktop computer

机译:从头细菌基因组测序:在台式计算机上组装的数百万个短读

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摘要

Novel high-throughput DNA sequencing technologies allow researchers to characterize a bacterial genome during a single experiment and at a moderate cost. However, the increase in sequencing throughput that is allowed by using such platforms is obtained at the expense of individual sequence read length, which must be assembled into longer contigs to be exploitable. This study focuses on the Illumina sequencing platform that produces millions of very short sequences that are 35 bases in length. We propose a de novo assembler software that is dedicated to process such data. Based on a classical overlap graph representation and on the detection of potentially spurious reads, our software generates a set of accurate contigs of several kilobases that cover most of the bacterial genome. The assembly results were validated by comparing data sets that were obtained experimentally for Staphylococcus aureus strain MW2 and Helicobacter acinonychis strain Sheeba with that of their published genomes acquired by conventional sequencing of 1.5- to 3.0-kb fragments. We also provide indications that the broad coverage achieved by high-throughput sequencing might allow for the detection of clonal polymorphisms in the set of DNA molecules being sequenced.
机译:新颖的高通量DNA测序技术使研究人员能够在单个实验中以中等成本表征细菌基因组。但是,使用此类平台所允许的测序通量的增加是以牺牲单个序列读取长度为代价的,必须将其组装成更长的重叠群才能利用。这项研究的重点是Illumina测序平台,该平台可产生数百万个长度为35个碱基的非常短的序列。我们建议使用从头汇编程序软件来处理此类数据。基于经典的重叠图表示法和对潜在的假读的检测,我们的软件可生成一组覆盖数千个碱基的准确碱基重叠群,覆盖大多数细菌基因组。通过比较通过实验获得的金黄色葡萄球菌菌株MW2和棘孢幽门螺杆菌Sheeba菌株的数据集与通过1.5到3.0-kb片段常规测序获得的已发表基因组的数据集,验证了组装结果。我们还提供了这样的迹象,即通过高通量测序获得的广泛覆盖范围可能允许检测正在测序的DNA分子集中的克隆多态性。

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