Forest soil fungal community elevational distribution pattern
Soil microbial DNA extraction was obtained and purified. ITS2 region of of soil fungi ribosome was amplified. The reverse primer combined with a barcode sequence. PCR amplification was performed in a 50 l reaction, containing 5 l 10Taq Buffer, 1.5 l dNTP Mixture, 0.5 l Taq, 2 l BSA (5mg/mL), 1 l primers, respectively, 1 l soil microbial template DNA (~ 25 ng/l) and 38 l ddH2O. The mixtures were amplified as following conditions: initial denaturation at 94 for 1 min, followed by 35 cycles of 94 for 20 s, 56 for 25 s and 68 for 45 s, with final extension at 68 for 10 min. The PCR products were purified and run using a Miseq (Illumina, San Diego, CA, USA).
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