Abstract

The reduced cost of high‐throughput sequencing and the development of gene sets with wide phylogenetic applicability has led to the rise of sequence capture methods as a plausible platform for both phylogenomics and population genomics in plants. An important consideration in large targeted sequencing projects is the per‐sample cost, which can be inflated when using off‐the‐shelf kits or reagents not purchased in bulk. Here, we discuss methods to reduce per‐sample costs in high‐throughput targeted sequencing projects. We review the minimal equipment and consumable requirements for targeted sequencing while comparing several alternatives to reduce bulk costs in DNA extraction, library preparation, target enrichment, and sequencing. We consider how each of the workflow alterations may be affected by DNA quality (e.g., fresh vs. herbarium tissue), genome size, and the phylogenetic scale of the project. We provide a cost calculator for researchers considering targeted sequencing to use when designing projects, and identify challenges for future development of low‐cost sequencing in non‐model plant systems.

Highlights

  • Strategies for reducing per-sample costs in target capture sequencing for phylogenomics and population genomics in plants

  • The reduced cost of high-throughput sequencing and the development of gene sets with wide phylogenetic applicability has led to the rise of sequence capture methods as a ­plausible platform for both phylogenomics and population genomics in plants

  • We review the minimal equipment and consumable requirements for targeted sequencing while comparing several alternatives to reduce bulk costs in DNA extraction, library preparation, target enrichment, and sequencing

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Summary

INVITED SPECIAL ARTICLE

For the Special Issue: Conducting Botanical Research with Limited Resources: Low-Cost Methods in the Plant Sciences. Strategies for reducing per-sample costs in target capture sequencing for phylogenomics and population genomics in plants. The reduced cost of high-throughput sequencing and the development of gene sets with wide phylogenetic applicability has led to the rise of sequence capture methods as a ­plausible platform for both phylogenomics and population genomics in plants. We discuss methods to reduce per-sample costs in high-throughput targeted sequencing projects. The most common design for target sequence capture in plants involves developing probes that target low-copy orthologous loci from a narrow set of in-group taxa (Gardner et al, 2016; Vatanparast et al, 2018; Villaverde et al, 2018; Soto Gomez et al, 2019).

Sequencing Overall
DNA EXTRACTION
PURIFICATION AND SIZE SELECTION
LIBRARY PREPARATION
HYBRIDIZATION AND ENRICHMENT
Findings
LITERATURE CITED
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