title: Fungi ITS sequencing of cinnamon soil
design: DNA extraction of cinnamon soil samples was carried out by Ezup column soil genomic DNA extraction kit (Shanghai Sanggong Bioengineering Co., Ltd.), the soil use was 100~300 mg of soil, the specific operation was carried out according to the instructions provided by the kit, and the obtained DNA solution was quantified by NanoDrop One/Onec ultra-micro spectrophotometer, and its integrity was detected by 1% agarose gel electrophoresis, and stored at -20 C or directly used for subsequent experiments. After extracting the genomic DNA of the brown soil microorganisms, fungal ITS was amplified by ITS1+ITS2 (F:5-CTTGGTCATTTAGAGGAAGTAA-3, R:5- GCTGCGTTCTTCATCGATGC-3) primers. The total PCR reaction system was as follows: 5 L of 10 buffer, 1.0 L of 2.5 mmol/L dNTPs, 2.5 L of 10 mol/L upstream and downstream primers, 1.0 L of 5 U/L Taq enzyme, 20 ng of DNA template, and 50 L of sterilized deionized water. The PCR reaction program was set to: 94 C denaturation for 3 min; Then 94C for 30 s, 55C for 30 s, 72C for 30 s, and the reaction was carried out for 30 cycles. Finally, extend at 72 C for 10 min. After the PCR products were detected by 1.5% agarose gel electrophoresis, the sequencing libraries were purified and homogenized, and the qualified libraries were sequenced with Illumina HiSeq PE250 and sequenced by Shanghai Sanggong Co., Ltd. Sample Barcode: CK1, ATCTGCA; U1, ATGCTAG; UA1, ATCGCAC; CK3, ATAGAGT; U3, AGATAGT; UA3, AGCATCG; CK5, AGTCGTC; U5, AGAGCAG; UA5, AGCTCTA; CK7, AGTATAC; U7, AGTGCGA; UA7, ACACACG; CK9, ACAGCGA; U9, ACATACT; UA9, ACTCTCA.