NODEOtherTing ZhuAMPLICONhttps://www.biosino.org/node/experiment/detail/OEX00002156Illumina HiSeq 2500RANDOMPaired564['human skin metagenome']NSA3 (5'-AAACTCTGTCGTGCTGGGGATA-3') and NLC2 (5'-GAGCTGCATTCCCAAACAACTC-3') were used for the first round of nested PCR. Each 25-µl reaction contained 12.5 µl Ex Taq Premix v.2.0 (Takara, Dalian, China), 1 µl each of the forward and reverse primers (2.5 µM), 5 µl template DNA, 1 µl bovine serum albumin (20 mg/ml), and 4.5 µl ddH2O. PCR reaction conditions were as follows: 94°C for 5 min; 30 cycles of 94°C for 30 s, 50°C for 45 s, and 72°C for 60 s; and 72°C for 10 min. NSI1 (5'-NNNNNNNNNNNNCA GATTGAATGGCTTAGTGAGG-3') and 58A2R (5'-NNNNNNNNNNNNCA CTGCGTTCTTCATCGAT-3') with 12-nt barcodes were used as primers for the second round. Second-round PCR was similar to the first round except that 2.5 µl of product from the first round was used as template and the number of amplification cycles was 25 instead of 30. PCR-amplified products were pooled and purified using the AxyPrep DNA Gel Extraction kit (Axygen, Tewksbury, MA, USA) according to the manufacturer’s instructions, then quantified with the dsDNA HS Assay (Invitrogen, Darmstadt, Germany) and used for library construction for Illumina sequencing with the LTP Library Preparation kit (KAPA Biosystems, Boston, MA, USA). Sequencing data were obtained with an Illumina Miseq platform (San Diego, CA, USA) using the 2*300 paired-end protocol.NODEfalseAmplicon libraries of ITS1 genesDNA was extracted within 24 hour using the MoBio PowerSoil DNA Isolation kit (Qiagen, Valencia, CA, USA). Nested PCR was used to amplify the fungal internal transcribed spacer 1 (ITS1) using barcoded-primers. PCR-amplified products were pooled and purified using the AxyPrep DNA Gel Extraction kit (Axygen, Tewksbury, MA, USA) according to the manufacturer’s instructions, then quantified with the dsDNA HS Assay (Invitrogen, Darmstadt, Germany) and used for library construction for Illumina sequencing with the LTP Library Preparation kit (KAPA Biosystems, Boston, MA, USA). Sequencing data were obtained with an Illumina Miseq platform (San Diego, CA, USA) using the 2*300 paired-end protocol.2020-03-242019-12-24OEX00002156OEP00000722