Diversity and abundance of fungi in bioaerosol from a gassy coal mine with methane by pyrosequencing and real time PCR
Nested PCR was performed for fungal communities. FUN-NS1/NS6R and FUN-V4F1/V4R2 were served as the first and second PCR amplification primers for fungi. The PCR reaction was initiated at 95oC for 2 min, followed by 25 cycles of 95oC for 30 sec, 55oC for 30 sec, 72oC for 60 sec, extended at 72oC for 5 min. PCR products were purified with the PCR purification kit and then prepared for pyrosequencing in Shanghai Majorbio Bio-pharm Biotechnology Company.