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

Tremella fuciformis

Gut & microbiomeSkin & hydration

SpeciesSnow Fungus, Silver Ear

JournalMycobiology

Year2019

Key points

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

Metadata-grounded summary

Citation 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.

Citation

Wang Y, Xu D, Liu D, Sun X, Chen Y, Zheng L, et al. (2019). A Rapid and Effective Colony PCR Procedure for Screening Transformants in Several Common Mushrooms. Mycobiology https://doi.org/10.1080/12298093.2019.1628523 PMID: 31565472

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