Cargando…
Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1)
• Premise of the study: As systematists grapple with how to best harness the power of next-generation sequencing (NGS), a deluge of review papers, methods, and analytical tools make choosing the right method difficult. Oreocarya (Boraginaceae), a genus of 63 species, is a good example of a group lac...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Botanical Society of America
2014
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259456/ https://www.ncbi.nlm.nih.gov/pubmed/25506521 http://dx.doi.org/10.3732/apps.1400062 |
_version_ | 1782348020325548032 |
---|---|
author | Ripma, Lee A. Simpson, Michael G. Hasenstab-Lehman, Kristen |
author_facet | Ripma, Lee A. Simpson, Michael G. Hasenstab-Lehman, Kristen |
author_sort | Ripma, Lee A. |
collection | PubMed |
description | • Premise of the study: As systematists grapple with how to best harness the power of next-generation sequencing (NGS), a deluge of review papers, methods, and analytical tools make choosing the right method difficult. Oreocarya (Boraginaceae), a genus of 63 species, is a good example of a group lacking both species-level resolution and genomic resources. The use of Geneious removes bioinformatic barriers and makes NGS genome skimming accessible to even the least tech-savvy systematists. • Methods: A combination of de novo and reference-guided assemblies was used to process 100-bp single-end Illumina HiSeq 2000 reads. A subset of 25 taxa was used to test the suitability of genome skimming for future systematic studies in recalcitrant lineages like Oreocarya. • Results: The nuclear ribosomal cistron, the plastome, and 12 mitochondrial genes were recovered from all 25 taxa. All data processing and phylogenomic analyses were performed in Geneious. We report possible future multiplexing levels and published low-copy nuclear genes represented within de novo contigs. • Discussion: Genome skimming represents a much-improved primary data collection over PCR+Sanger sequencing when chloroplast DNA (cpDNA), nuclear ribosomal DNA (nrDNA), and mitochondrial DNA (mtDNA) are the target sequences. This study details methods that plant systematists can employ to study their own taxa of interest. |
format | Online Article Text |
id | pubmed-4259456 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Botanical Society of America |
record_format | MEDLINE/PubMed |
spelling | pubmed-42594562014-12-12 Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) Ripma, Lee A. Simpson, Michael G. Hasenstab-Lehman, Kristen Appl Plant Sci Application Article • Premise of the study: As systematists grapple with how to best harness the power of next-generation sequencing (NGS), a deluge of review papers, methods, and analytical tools make choosing the right method difficult. Oreocarya (Boraginaceae), a genus of 63 species, is a good example of a group lacking both species-level resolution and genomic resources. The use of Geneious removes bioinformatic barriers and makes NGS genome skimming accessible to even the least tech-savvy systematists. • Methods: A combination of de novo and reference-guided assemblies was used to process 100-bp single-end Illumina HiSeq 2000 reads. A subset of 25 taxa was used to test the suitability of genome skimming for future systematic studies in recalcitrant lineages like Oreocarya. • Results: The nuclear ribosomal cistron, the plastome, and 12 mitochondrial genes were recovered from all 25 taxa. All data processing and phylogenomic analyses were performed in Geneious. We report possible future multiplexing levels and published low-copy nuclear genes represented within de novo contigs. • Discussion: Genome skimming represents a much-improved primary data collection over PCR+Sanger sequencing when chloroplast DNA (cpDNA), nuclear ribosomal DNA (nrDNA), and mitochondrial DNA (mtDNA) are the target sequences. This study details methods that plant systematists can employ to study their own taxa of interest. Botanical Society of America 2014-12-04 /pmc/articles/PMC4259456/ /pubmed/25506521 http://dx.doi.org/10.3732/apps.1400062 Text en © 2014 Ripma et al. Published by the Botanical Society of America http://creativecommons.org/licenses/by-nc/4.0/ This work is licensed under a Creative Commons Attribution License (CC-BY-NC-SA). |
spellingShingle | Application Article Ripma, Lee A. Simpson, Michael G. Hasenstab-Lehman, Kristen Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title | Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title_full | Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title_fullStr | Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title_full_unstemmed | Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title_short | Geneious! Simplified genome skimming methods for phylogenetic systematic studies: A case study in Oreocarya (Boraginaceae)(1) |
title_sort | geneious! simplified genome skimming methods for phylogenetic systematic studies: a case study in oreocarya (boraginaceae)(1) |
topic | Application Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4259456/ https://www.ncbi.nlm.nih.gov/pubmed/25506521 http://dx.doi.org/10.3732/apps.1400062 |
work_keys_str_mv | AT ripmaleea geneioussimplifiedgenomeskimmingmethodsforphylogeneticsystematicstudiesacasestudyinoreocaryaboraginaceae1 AT simpsonmichaelg geneioussimplifiedgenomeskimmingmethodsforphylogeneticsystematicstudiesacasestudyinoreocaryaboraginaceae1 AT hasenstablehmankristen geneioussimplifiedgenomeskimmingmethodsforphylogeneticsystematicstudiesacasestudyinoreocaryaboraginaceae1 |