eDNA from waters where no mangroves grow, south to Marsa Alam City
Environmental DNA (eDNA) was extracted from membrane filters belonging three sampling points in this non-mangrove site using a commercial silica gel column-based kit under sterile conditions. DNA from the sampling points in this site was pooled, quantified via UV absorbance, and adjusted for purity before PCR amplification of the COI gene using indexed primers. PCR products were verified by gel electrophoresis. Then, PCR product from this site was pooled with that from the other sites, based on site-specific index primers. Then, library preparation followed Illuminas 16S protocol yet using COI primers instead of the 16S ones. Final libraries were cleaned, quantified, and sequenced on the MiSeq platform using the Reagent Kit v3.