16S rRNA of Rhizospheric Microorganism:Tamarind
This study analyzed the diversity of the microbial 16S rRNA V3V4 region. Samples (0.2g) were ground with lysis buffer using a Tissuelyser-48 (60Hz), and DNA was extracted via MagBeads FastDNA Kit, quantified by Nanodrop, and verified by 0.8% agarose gel. The V3V4 region was amplified with barcoded primers 338F/806R using ABclonal polymerase (98C 5min; 26 cycles: 98C 30s, 52C 30s, 72C 45s; 72C 5min). Amplicons were gel-recovered (2%) and libraries constructed with TruSeq Nano Kit (end repair, A-tailing, adapter ligation, PCR). Fragments were size-selected (2% gel), purified by beads, quantified by PicoGreen/Agilent Bioanalyzer, and sequenced on NovaSeq 6000 (2250bp). Data were analyzed via QIIME2 2022.11: demux, cutadapt, DADA2 for ASVs, Greengenes2 annotation, and diversity analyses (Alpha/Beta diversity, PICRUSt14 function).
SUB15408876
TG94-R