Amplicon-Metagenomic Analysis of Phlogacanthus thyrsiformis
The leaves were subjected to surface sterilization. Briefly, the leaves were washed with running tap water and sterile distilled water for 60 s each followed by washing with 5% sodium hypochlorite (NaOCl) for 5 min and finally rinsing with distilled water at least three times (Lacerda et al. 2018). The leaves were ready for the extraction of DNA (for amplicon metagenome sequencing). Nearly, 500 mg of each healthy surface sterilized leaves was used for the DNA extraction using FastDNA SPIN Kit for Soil (SKU:116560200, MP Biomedicals, Santa Ana, California, USA) following instructions by manufacturer. In the study, sample Phlogacanthus tubiflorus will be referred to as SKS_1, sample P. thyrsiformis will be referred as SKS_2, and sample P. curviflorus was referred as SKS_3. The quality and quantity of DNA were assessed using both Nanodrop Spectrophotometer and Qubit DNA HS Assay Kit (Catalog No. Q32851) (Invitrogen, Waltham, Massachusetts, U.S.) and gel electrophoresis was used for assessing the condition of the DNA.