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A novel method of genomic DNA extraction for Cactaceae(1)

• Premise of the study: Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated. • Methods and Results: Several protocols...

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Detalles Bibliográficos
Autores principales: Fehlberg, Shannon D., Allen, Jessica M., Church, Kathleen
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Botanical Society of America 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4105278/
https://www.ncbi.nlm.nih.gov/pubmed/25202521
http://dx.doi.org/10.3732/apps.1200013
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author Fehlberg, Shannon D.
Allen, Jessica M.
Church, Kathleen
author_facet Fehlberg, Shannon D.
Allen, Jessica M.
Church, Kathleen
author_sort Fehlberg, Shannon D.
collection PubMed
description • Premise of the study: Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated. • Methods and Results: Several protocols for extracting DNA from spines were tested and modified to maximize yield, amplification, and sequencing. Sampling of and extraction from spines resulted in a simplified protocol overall and complete avoidance of mucilage as compared to typical tissue extractions. Sequences from one nuclear and three plastid regions were obtained across eight genera and 20 species of cacti using DNA extracted from spines. • Conclusions: Genomic DNA useful for amplification and sequencing can be obtained from cactus spines. The protocols described here are valuable for any cactus species, but are particularly useful for investigators interested in sampling living collections, extensive field sampling, and/or conservation genetic studies.
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spelling pubmed-41052782014-09-08 A novel method of genomic DNA extraction for Cactaceae(1) Fehlberg, Shannon D. Allen, Jessica M. Church, Kathleen Appl Plant Sci Protocol Note • Premise of the study: Genetic studies of Cactaceae can at times be impeded by difficult sampling logistics and/or high mucilage content in tissues. Simplifying sampling and DNA isolation through the use of cactus spines has not previously been investigated. • Methods and Results: Several protocols for extracting DNA from spines were tested and modified to maximize yield, amplification, and sequencing. Sampling of and extraction from spines resulted in a simplified protocol overall and complete avoidance of mucilage as compared to typical tissue extractions. Sequences from one nuclear and three plastid regions were obtained across eight genera and 20 species of cacti using DNA extracted from spines. • Conclusions: Genomic DNA useful for amplification and sequencing can be obtained from cactus spines. The protocols described here are valuable for any cactus species, but are particularly useful for investigators interested in sampling living collections, extensive field sampling, and/or conservation genetic studies. Botanical Society of America 2013-03-05 /pmc/articles/PMC4105278/ /pubmed/25202521 http://dx.doi.org/10.3732/apps.1200013 Text en © 2013 Fehlberg 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 Protocol Note
Fehlberg, Shannon D.
Allen, Jessica M.
Church, Kathleen
A novel method of genomic DNA extraction for Cactaceae(1)
title A novel method of genomic DNA extraction for Cactaceae(1)
title_full A novel method of genomic DNA extraction for Cactaceae(1)
title_fullStr A novel method of genomic DNA extraction for Cactaceae(1)
title_full_unstemmed A novel method of genomic DNA extraction for Cactaceae(1)
title_short A novel method of genomic DNA extraction for Cactaceae(1)
title_sort novel method of genomic dna extraction for cactaceae(1)
topic Protocol Note
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4105278/
https://www.ncbi.nlm.nih.gov/pubmed/25202521
http://dx.doi.org/10.3732/apps.1200013
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