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

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