title: Bacterial 16S 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, bacterial 16S was amplified using V4+V5 (F:5-ACTCCTACGGGAGGCAGCAG-3, R: 5-GGACTACHVGGGTWTCTAAT-3). 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, GGGATT; U1, TATTTC; UA1, GAAAGA; CK3, CGTAAA; U3, CTTTGG; UA3, TACTTT; CK5, CCCGGT; U5, TGCAAA; UA5,TACCCT; CK7, CCCTTT; U7, CGGGCT; UA7, CCCTGG; CK9, CGGGAA; U9, AGAAAG; UA9, TTTGTC;