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利用读长深度、k-mer和流式细胞术测量基因组大小:甲虫(鞘翅目)的方法学比较

Measuring Genome Sizes Using Read-Depth, k-mers, and Flow Cytometry: Methodological Comparisons in Beetles (Coleoptera).

作者信息

Pflug James M, Holmes Valerie Renee, Burrus Crystal, Johnston J Spencer, Maddison David R

机构信息

Department of Integrative Biology, Oregon State University, Corvallis, OR 97331

Department of Entomology, Texas A&M University, College Station, TX 77843.

出版信息

G3 (Bethesda). 2020 Sep 2;10(9):3047-3060. doi: 10.1534/g3.120.401028.

Abstract

Measuring genome size across different species can yield important insights into evolution of the genome and allow for more informed decisions when designing next-generation genomic sequencing projects. New techniques for estimating genome size using shallow genomic sequence data have emerged which have the potential to augment our knowledge of genome sizes, yet these methods have only been used in a limited number of empirical studies. In this project, we compare estimation methods using next-generation sequencing (k-mer methods and average read depth of single-copy genes) to measurements from flow cytometry, a standard method for genome size measures, using ground beetles (Carabidae) and other members of the beetle suborder Adephaga as our test system. We also present a new protocol for using read-depth of single-copy genes to estimate genome size. Additionally, we report flow cytometry measurements for five previously unmeasured carabid species, as well as 21 new draft genomes and six new draft transcriptomes across eight species of adephagan beetles. No single sequence-based method performed well on all species, and all tended to underestimate the genome sizes, although only slightly in most samples. For one species, sp. nr. , most sequence-based methods yielded estimates half the size suggested by flow cytometry.

摘要

测量不同物种的基因组大小能够为基因组进化提供重要见解,并且在设计下一代基因组测序项目时有助于做出更明智的决策。利用浅层基因组序列数据估算基因组大小的新技术已经出现,这些技术有可能扩充我们对基因组大小的认识,但这些方法仅在有限数量的实证研究中得到应用。在本项目中,我们将使用下一代测序的估算方法(k-mer方法和单拷贝基因的平均读长深度)与流式细胞术测量结果进行比较,流式细胞术是测量基因组大小的标准方法,我们以步甲科昆虫(步甲科)和肉食亚目甲虫的其他成员作为测试系统。我们还提出了一种利用单拷贝基因读长深度估算基因组大小的新方案。此外,我们报告了五种此前未测量过的步甲物种的流式细胞术测量结果,以及八种肉食亚目甲虫的21个新基因组草图和六个新转录组草图。没有一种基于序列的方法在所有物种上都表现良好,所有方法都倾向于低估基因组大小,不过在大多数样本中低估幅度较小。对于一个物种,sp. nr.,大多数基于序列的方法得出的估计值仅为流式细胞术所显示大小的一半。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/f57b/7466995/d5095639c1f4/3047f1.jpg

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