AI Article Synopsis
- Third-generation sequencing allows for longer DNA reads, enhancing studies in biodiversity and genetics.
- We developed a cost-effective hybrid capture protocol to enrich and capture long plastid genome fragments from six different plant species.
- The method yielded high enrichment rates for plastome fragments, though DNA from dried leaves produced lower quality results compared to fresh leaves.
Article Abstract
Premise: Third-generation sequencing methods generate significantly longer reads than those produced using alternative sequencing methods. This provides increased possibilities for the study of biodiversity, phylogeography, and population genetics. We developed a protocol for in-solution enrichment hybridization capture of long DNA fragments applicable to complete plastid genomes.
Methods And Results: The protocol uses cost-effective in-house probes developed via long-range PCR and was used in six non-model monocot species (Poaceae: African rice, pearl millet, fonio; and three palm species). DNA was extracted from fresh and silica gel-dried leaves. Our protocol successfully captured long-read plastome fragments (3151 bp median on average), with an enrichment rate ranging from 15% to 98%. DNA extracted from silica gel-dried leaves led to low-quality plastome assemblies when compared to DNA extracted from fresh tissue.
Conclusions: Our protocol could also be generalized to capture long sequences from specific nuclear fragments.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6526642 | PMC |
| http://dx.doi.org/10.1002/aps3.1243 | DOI Listing |
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