16S rRNA metagenome sequencing of earthworm gut
The sample was divided into two parts. First part of the sample was centrifuged at 15,000 xg for 2 minutes. Supernatant was discarded. CD1 was added to the (250mg) samples and heated at 650C for 10 minutes and proceeded as per manufacturers instruction using the Dneasy PowerSoil Pro Kit (Cat No./ID: 47014, QIAGEN GmbH) as instructed by manufacturer. The DNA were quantified on NanoDropOne and Qubit using water and standard as a control respectively. Libraries were constructed in alignment withthe 16S metagenomic library preparation protocol from Illumina Inc. The genespecific sequencesused in this protocol target the 16S V3 and V4 region. They are selected from the Klindworth et al., as the most promising bacterial primer pair. Illumina adapter overhang nucleotide sequences are added to the genespecific sequences.