KMID : 1160220190470030350
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Mycobiology 2019 Volume.47 No. 3 p.350 ~ p.354
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A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms
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Wang Yuanyuan
Xu Danyun Liu Dongmei Sun Xueyan Chen Yue Zheng Lisheng Chen Liguo Ma Aimin
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Abstract
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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?¥ìg) to 20?¥ìl 0.25% Lywallzyme solution, and vortexing for 10?s followed by incubation at 34?¡ÆC for 15?min. Finally, 2?¥ìl 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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KEYWORD
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Colony PCR, mushroom, screening, transformant
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