A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms

一种快速有效的菌落 PCR 方法,用于筛选几种常见蘑菇中的转化子

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作者:Yuanyuan Wang, Danyun Xu, Dongmei Liu, Xueyan Sun, Yue Chen, Lisheng Zheng, Liguo Chen, Aimin Ma

Abstract

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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