Microbial community analysis via 16S rDNA
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Corresponding Organization :
Other organizations : Shandong University
Variable analysis
- Primers 338F (ACTCCTACGGGAGGCAGCAG) and 806R (GGACTACHVGGGTWTCTAAT)
- 16S rDNA amplification in the V3–V4 region
- Genomic DNA extraction using the genome extraction kit (TIANGEN BIOTECH, China)
- PCR conditions: initial denaturation at 95 °C for 3 min; 30 cycles of denaturation at 95 °C for 10 s, annealing at 56 °C for 15 s, and extension at 72 °C for 1.5 min; then final extension at 72 °C for 10 min
- Phanta Max Super-Fidelity DNA Polymerase (Vazyme, China) used for PCR amplification
- Expected length of the PCR product was 1.6 kb
- Amplicon sequencing using the NovaSeq platform (Allwegene, China)
- At least 40,000 reads acquired for each sample
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