Brassica campestris L. soil rhizosphere microecology
收藏科学数据银行2025-12-05 更新2026-04-23 收录
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Nucleic acid was extracted from rhizosphere soil samples using the MagBeams FastDNA Kit for Soil assay kit; For the extracted DNA, the molecular size was judged by 0.8% agarose gel electrophoresis, and the DNA was quantified by Nanodrop. Select the highly variable V3-V4 regions of the bacterial 16S rRNA gene for sequencing. Primers used for PCR amplification: 338F (5 '- ACTCCTACGGGAGGCAGCA-3'), 806R (5 '- GACTACHVGGGTWTCTAAT-3'). Quantify PCR products using Quant iT PicoGreen dsDNA Assay Kit, then mix samples according to the required data volume for each sample, and build libraries using Illumina's TruSeq Nano DNA LT Library Prep Kit. Perform 2 × 250bp double ended sequencing using NovaSeq 6000 SP Reagent Kit (500 cycles) on Illumina NovaSeq machine. The original sequence data is decoded using the Demux plugin, primer excision is performed using the cutadapt plugin, and then the sequence is processed using the DADA2 plugin for quality filtering, denoising, concatenation, and chimerism removal. Merge the obtained sequences with 100% sequence similarity to generate characteristic sequences ASVs and abundance data tables.
提供机构：
li qian
创建时间：
2025-12-05
