An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex
Abstract Premise The isolation of RNA from trees is challenging due to the interference of polyphenols and polysaccharides with downstream processes. Furthermore, many RNA extraction protocols are time consuming and involve hazardous chemicals. To address these issues, we aimed to develop a safe pro...
| Main Authors: | , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-05-01
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| Series: | Applications in Plant Sciences |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/aps3.11518 |
| _version_ | 1797800182072475648 |
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| author | Maryam Hadi Elizabeth A. Stacy |
| author_facet | Maryam Hadi Elizabeth A. Stacy |
| author_sort | Maryam Hadi |
| collection | DOAJ |
| description | Abstract Premise The isolation of RNA from trees is challenging due to the interference of polyphenols and polysaccharides with downstream processes. Furthermore, many RNA extraction protocols are time consuming and involve hazardous chemicals. To address these issues, we aimed to develop a safe protocol for high‐quality RNA extraction from diverse Metrosideros taxa representing a broad range of leaf toughness, pubescence, and secondary metabolites. Methods and Results We tested popular RNA isolation kits and protocols that were effective on other recalcitrant trees, including a broad range of optimization and purification steps. We optimized a protocol involving two silica‐membrane column‐based kits that yielded high‐quantity RNA with an RNA integrity number >7 and without DNA contamination. All RNA samples were used successfully in a follow‐on RNA‐Seq experiment. Conclusions We present an optimized high‐throughput RNA extraction protocol that yielded high‐quality and high‐quantity RNA from three contrasting leaf phenotypes within a hyperdiverse woody species complex. |
| first_indexed | 2024-03-13T04:30:12Z |
| format | Article |
| id | doaj.art-ae857ea5e546415690e60b1ede63fe58 |
| institution | Directory Open Access Journal |
| issn | 2168-0450 |
| language | English |
| last_indexed | 2024-03-13T04:30:12Z |
| publishDate | 2023-05-01 |
| publisher | Wiley |
| record_format | Article |
| series | Applications in Plant Sciences |
| spelling | doaj.art-ae857ea5e546415690e60b1ede63fe582023-06-19T16:55:29ZengWileyApplications in Plant Sciences2168-04502023-05-01113n/an/a10.1002/aps3.11518An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complexMaryam Hadi0Elizabeth A. Stacy1School of Life Sciences University of Nevada Las Vegas Las Vegas Nevada 89154 USASchool of Life Sciences University of Nevada Las Vegas Las Vegas Nevada 89154 USAAbstract Premise The isolation of RNA from trees is challenging due to the interference of polyphenols and polysaccharides with downstream processes. Furthermore, many RNA extraction protocols are time consuming and involve hazardous chemicals. To address these issues, we aimed to develop a safe protocol for high‐quality RNA extraction from diverse Metrosideros taxa representing a broad range of leaf toughness, pubescence, and secondary metabolites. Methods and Results We tested popular RNA isolation kits and protocols that were effective on other recalcitrant trees, including a broad range of optimization and purification steps. We optimized a protocol involving two silica‐membrane column‐based kits that yielded high‐quantity RNA with an RNA integrity number >7 and without DNA contamination. All RNA samples were used successfully in a follow‐on RNA‐Seq experiment. Conclusions We present an optimized high‐throughput RNA extraction protocol that yielded high‐quality and high‐quantity RNA from three contrasting leaf phenotypes within a hyperdiverse woody species complex.https://doi.org/10.1002/aps3.11518Metrosiderosprotocol developmentRNA extractiontreeswoody species |
| spellingShingle | Maryam Hadi Elizabeth A. Stacy An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex Applications in Plant Sciences Metrosideros protocol development RNA extraction trees woody species |
| title | An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex |
| title_full | An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex |
| title_fullStr | An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex |
| title_full_unstemmed | An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex |
| title_short | An optimized RNA extraction method for diverse leaves of Hawaiian Metrosideros, a hypervariable tree species complex |
| title_sort | optimized rna extraction method for diverse leaves of hawaiian metrosideros a hypervariable tree species complex |
| topic | Metrosideros protocol development RNA extraction trees woody species |
| url | https://doi.org/10.1002/aps3.11518 |
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