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Once a single fungal colony appears, it can promptly spread throughout the medium plate and interfere with screening.
After appropriate dilutions in the buffer solution, aliquots of 100 μL were spread onto an agar medium plate and incubated at 37°C for 15 h.
The cells were re-seeded (750 cells per medium plate) and incubated for 10 days.
A single colony was picked from each of the selection medium plate and β Galactosidase assay was performed as described in the methods section.
The seedlings were then transferred back to the growth medium plate and incubated in either continuous light or darkness for various length of time before being subjected to laser confocal scanning microscopic observation.
Approximately 0.1 mL of each of the other samples was placed directly onto a 7H10 Middlebrook medium plate and spread over the plate by using a sterile loop.
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After incubation 5 µl from each wells were inoculated on agar medium plates and the plates were incubated for 24 h.
For spore production, the fungus was grown on potato dextrose agar (PDA) medium plates and incubated at 37 °C for 72 h.
For charcoal mating assays, strains were crossed on charcoal-containing complete medium plates and incubated at 22°C [46].
After 14 days, seedlings with long or short roots were individually transferred to MS medium plates and 2 3 leaves were removed to prepare DNA for PCR analysis.
Two single knock-out strains were crossed on BMM medium plates and asci from recombinant perithecia were isolated after 7 10 days.
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