Soil Consortia 1
Nucleospin soil kit (MN) was used to extract DNA following the manufacturers instructions. V3-V4 amplicons were generated directly from the DNA using a nested PCR strategy. The Kapa Hyperplus kit was used to perform library prep. The generated amplicon was purified followed by end repair and A-tailing to create end-repaired, 5'-phosphorylated, 3'-dA-tailed dsDNA fragments. Subsequently, adapter ligation was performed, where dsDNA adapters with 3'-dTMP overhangs were ligated to the 3'-dA-tailed fragments. The library generated was then amplified with minimal PCR cycles to avoid any bias. The libraries were normalized, pooled and sequenced using MiSeq. V3 500 cycle kit was used to generate 0.1 million reads per sample with 250bp read length.