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A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field

A major challenge in extracting high-quality DNA from bryophytes is the treatment of bryophyte material in the field. The existing and commonly used treatment methods in the field have several shortcomings. Natural drying methods can lead to DNA breaks. In addition, it is highly cumbersome to carry...

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Autores principales: Shen, Fengjiao, Li, Lin, Wang, Dan, Wang, Mengzhen, Shevock, James R., Zhao, Jiancheng, Shi, Shuo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9683613/
https://www.ncbi.nlm.nih.gov/pubmed/36417395
http://dx.doi.org/10.1371/journal.pone.0277778
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author Shen, Fengjiao
Li, Lin
Wang, Dan
Wang, Mengzhen
Shevock, James R.
Zhao, Jiancheng
Shi, Shuo
author_facet Shen, Fengjiao
Li, Lin
Wang, Dan
Wang, Mengzhen
Shevock, James R.
Zhao, Jiancheng
Shi, Shuo
author_sort Shen, Fengjiao
collection PubMed
description A major challenge in extracting high-quality DNA from bryophytes is the treatment of bryophyte material in the field. The existing and commonly used treatment methods in the field have several shortcomings. Natural drying methods can lead to DNA breaks. In addition, it is highly cumbersome to carry large quantities of silica gel in the field due to its weight and high risk of contamination among samples. In this study, we explored more convenient drying methods to treat bryophyte specimens and promote more efficient DNA recovery. The quantity and quality of genomic DNA extracted from every bryophyte species using different drying methods, including hot-air drying methods (150°C, 80°C, and 40°C), natural drying method, and silica gel drying method, were measured. Spectrophotometry, electrophoresis, and PCR amplification were performed to assess the effects of different drying methods. The results of total DNA purity, total DNA concentration, PCR success, and OD 260/230 ratios suggested that the hot-air drying (40–80°C) was the best method. The morphological comparison revealed that hot-air drying at 40°C and 80°C exerted no significant adverse effects on plant morphology and taxonomic studies. Thus, this method prevents rapid DNA degradation and silica gel pollution and saves the workforce from carrying large amounts of silica gel to the field. Several inexpensive devices, such as portable hairdryers, fan heaters, and electric blankets, are available that can be easily carried to the field for drying molecular specimens.
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spelling pubmed-96836132022-11-24 A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field Shen, Fengjiao Li, Lin Wang, Dan Wang, Mengzhen Shevock, James R. Zhao, Jiancheng Shi, Shuo PLoS One Research Article A major challenge in extracting high-quality DNA from bryophytes is the treatment of bryophyte material in the field. The existing and commonly used treatment methods in the field have several shortcomings. Natural drying methods can lead to DNA breaks. In addition, it is highly cumbersome to carry large quantities of silica gel in the field due to its weight and high risk of contamination among samples. In this study, we explored more convenient drying methods to treat bryophyte specimens and promote more efficient DNA recovery. The quantity and quality of genomic DNA extracted from every bryophyte species using different drying methods, including hot-air drying methods (150°C, 80°C, and 40°C), natural drying method, and silica gel drying method, were measured. Spectrophotometry, electrophoresis, and PCR amplification were performed to assess the effects of different drying methods. The results of total DNA purity, total DNA concentration, PCR success, and OD 260/230 ratios suggested that the hot-air drying (40–80°C) was the best method. The morphological comparison revealed that hot-air drying at 40°C and 80°C exerted no significant adverse effects on plant morphology and taxonomic studies. Thus, this method prevents rapid DNA degradation and silica gel pollution and saves the workforce from carrying large amounts of silica gel to the field. Several inexpensive devices, such as portable hairdryers, fan heaters, and electric blankets, are available that can be easily carried to the field for drying molecular specimens. Public Library of Science 2022-11-23 /pmc/articles/PMC9683613/ /pubmed/36417395 http://dx.doi.org/10.1371/journal.pone.0277778 Text en © 2022 Shen et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Shen, Fengjiao
Li, Lin
Wang, Dan
Wang, Mengzhen
Shevock, James R.
Zhao, Jiancheng
Shi, Shuo
A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title_full A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title_fullStr A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title_full_unstemmed A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title_short A comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
title_sort comparison of drying methods on the quality for bryophyte molecular specimens collected in the field
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9683613/
https://www.ncbi.nlm.nih.gov/pubmed/36417395
http://dx.doi.org/10.1371/journal.pone.0277778
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