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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...

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Detalles Bibliográficos
Autores principales: Wang, Yuanyuan, Xu, Danyun, Liu, Dongmei, Sun, Xueyan, Chen, Yue, Zheng, Lisheng, Chen, Liguo, Ma, Aimin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2019
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.
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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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