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在DNBSEQ数据中分析软件读取交叉污染.

Dmitry N Konanov1, Vera Y Tereshchuk2, Ignat V Sonets1,3

  • 1Research Institute for System Biology and Medicine, Moscow 117246, Russia.

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概括

DNA纳米球测序 (DNBSEQ) PE300套件可以产生高数据,但可能会导致"软件污染"文物. 这些问题,如不当的阅读配对,发生在图书馆具有不同的插入大小或加载时,影响基因组和转录组分析.

关键词:
在 DNBSEQQ 中.数据过数据过.阅读复制件的复制件对文物进行测序.

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科学领域:

  • 基因组学就是基因组学.
  • 下一代测序的测序方法
  • 生物信息学是一种生物信息学.

背景情况:

  • DNA纳米球测序 (DNBSEQ) 是一个快速发展的测序技术.
  • 针对DNBSEQ-G99和G400的新PE300套件提供高数据产量,适用于基因组和转录基因组研究.
  • 在单次运行中将多种DNA库结合起来,可以引入数据工件.

研究的目的:

  • 调查在DNBSEQ PE300运行中观察到的"软件污染"文物的原因.
  • 描述这些文物的性质,包括读取配对和解复杂错误.
  • 评估这些文物对基因组和转录基因组数据分析的影响.

主要方法:

  • 对DNBSEQ PE300测序数据的分析,包括MGI演示数据集.
  • 检查阅读配对,解复和模拟阅读生成.
  • 将DNBSEQ数据与使用NA12878人类外体数据的Illumina测序数据进行比较.

主要成果:

  • 在所有分析的DNBSEQ运行中观察到"软件污染"的文物,例如不当的读取配对和解复,在所有分析的DNBSEQ运行中观察到.
  • 这些文物源于对来自邻近DNA纳米球的信号的误解,特别是在短插入序列中.
  • 在DNBSEQ数据中的不当配对率与Illumina相似,问题来自不同的插入尺寸分布或流量细胞负载.
  • 原始DNBSEQ数据可能包含大约2%的光学重复,因为DNA纳米球的距离很近.

结论:

  • 在DNBSEQ PE300测序中的工件主要是由相邻的DNA纳米球之间的信号干扰引起的.
  • 这些工件在测序具有不同插入大小分布或流细胞加载的库时显著影响数据质量.
  • 虽然不恰当的配对率与Illumina相似,但仔细的图书馆准备和数据分析对于使用DNBSEQ的精确基因组和转录组结果至关重要.