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

摘要

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.