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"通过全基因组牛津纳米孔测序评估多重位移放大效应的好处和局限性"

Fiifi A Dadzie¹, Megan S Beaudry², Alex Deyanov³

¹Department of Genetics, University of Georgia, Athens, GA USA 30602.

bioRxiv : the preprint server for biology

|February 26, 2024

概括

多重位移放大 (MDA) 能够从最小的DNA进行高质量的基因组测序. 一个新的工具CADECT有效地删除了文物序列,以改善基因组组装,特别是难以培养的生物体.

科学领域:

基因组学就是基因组学.
分子生物学分子生物学
生物信息学是一种生物信息学.

背景情况:

多重位移放大 (MDA) 在放大长DNA片段和整个基因组方面优于PCR.
对于测序有限的DNA样本来改善基因组组件,MDA非常有价值.
牛津纳米孔技术 (ONT) 提供长读序列,适合MDA生成的DNA.

研究的目的:

通过ONT测序来评估MDA对基因组组装的有效性和局限性.
以低DNA输入量来评估MDA的性能.
开发一种用于处理MDA相关文物的计算工具.

主要方法:

利用MDA从各种细菌和寄生虫样本进行全基因组放大.
雇佣了ONT快速图书馆准备和MinION排序.
开发并应用了合体检测工具 (CADECT) 管道,以识别和删除合体序列.

主要成果:

从只有0.025 ng的DNA样本中获得了近乎完整的基因组序列.
在MDA放大剪切DNA中观察到尺寸增大模式,与放大长片段和形成结合体有关.
CADECT成功地减轻了结合体的影响,使得连续基因组组装甚至从退化的DNA.

结论:

对于从有限的DNA量中生成高质量的基因组组件,MDA非常有效.
通过解决 concatemers,CADECT 管道显著改善了基因组组装连接性.
这些发现对于研究不能培养的生物和加速用稀缺的DNA样本进行临床诊断至关重要.

关键词:

它们是Apicomplexa的复合物.进入细菌体中的细菌.科科科科 (Enterococcaceae) 是一个植物.传染性疾病传染性疾病 LRS 特别版特别版低丰富度的DNA是什么

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