A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms
10.1080/12298093.2019.1628523
- Author:
Yuanyuan WANG
1
;
Danyun XU
;
Dongmei LIU
;
Xueyan SUN
;
Yue CHEN
;
Lisheng ZHENG
;
Liguo CHEN
;
Aimin MA
Author Information
1. College of Food Science and Technology, Huazhong Agricultural University, Wuhan, China. aiminma@mail.hzau.edu.cn
- Publication Type:Note
- From:Mycobiology
2019;47(3):350-354
- CountryRepublic of Korea
- Language:English
-
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.
