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A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms
In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: pic...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2019
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6758613/ https://www.ncbi.nlm.nih.gov/pubmed/31565472 http://dx.doi.org/10.1080/12298093.2019.1628523 |
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author | Wang, Yuanyuan Xu, Danyun Liu, Dongmei Sun, Xueyan Chen, Yue Zheng, Lisheng Chen, Liguo Ma, Aimin |
author_facet | Wang, Yuanyuan Xu, Danyun Liu, Dongmei Sun, Xueyan Chen, Yue Zheng, Lisheng Chen, Liguo Ma, Aimin |
author_sort | Wang, Yuanyuan |
collection | PubMed |
description | In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant’s tissue (1–10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments. |
format | Online Article Text |
id | pubmed-6758613 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-67586132019-09-27 A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms Wang, Yuanyuan Xu, Danyun Liu, Dongmei Sun, Xueyan Chen, Yue Zheng, Lisheng Chen, Liguo Ma, Aimin Mycobiology Research Note In the post-genomic era, gene function analysis has attracted much attention. Transformation is often needed to investigate gene function. In this study, an easy, rapid, reliable, and cost-effective colony polymerase chain reaction (PCR) method for screening mushroom transformants was developed: picking up a suitable amount of transformant’s tissue (1–10 μg) to 20 μl 0.25% Lywallzyme solution, and vortexing for 10 s followed by incubation at 34 °C for 15 min. Finally, 2 μl of the suspension was used as templates to perform PCR and single target bands were successfully amplified from respective transformants of Tremella fuciformis, Pleurotus ostreatus, and Pleurotus tuber-regium. This procedure could be widely employed for screening transformants in mushroom transformation experiments. Taylor & Francis 2019-06-22 /pmc/articles/PMC6758613/ /pubmed/31565472 http://dx.doi.org/10.1080/12298093.2019.1628523 Text en © 2019 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group on behalf of the Korean Society of Mycology. http://creativecommons.org/licenses/by-nc/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Research Note Wang, Yuanyuan Xu, Danyun Liu, Dongmei Sun, Xueyan Chen, Yue Zheng, Lisheng Chen, Liguo Ma, Aimin A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title | A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title_full | A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title_fullStr | A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title_full_unstemmed | A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title_short | A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms |
title_sort | rapid and effective colony pcr procedure for screening transformants in several common mushrooms |
topic | Research Note |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6758613/ https://www.ncbi.nlm.nih.gov/pubmed/31565472 http://dx.doi.org/10.1080/12298093.2019.1628523 |
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