Mycobiology.  2019 Sep;47(3):350-354. 10.1080/12298093.2019.1628523.

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

• Wang Y
• Xu D
• Liu D
• Sun X
• Chen Y
• Zheng L
• Chen L
• Ma A

• KMID: 2470148
• DOI: http://doi.org/10.1080/12298093.2019.1628523

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.

Keyword

Colony PCR; mushroom; screening; transformant

MeSH Terms

Agaricales*
Mass Screening*
Methods
Pleurotus
Polymerase Chain Reaction*