๐Ÿ’ง Water collection

โ†’ Collect environmental water samples using sterile techniques.

๐Ÿงช Filtration

โ†’ Pass water through membrane filters to capture suspended DNA (eDNA) from the environment.

๐Ÿงฌ DNA extraction

โ†’ Isolate DNA from the filters using a commercial extraction kit (e.g. Qiagen DNeasy).

๐Ÿ”ฌ DNA quantification (Qubit)

โ†’ Measure DNA concentration to assess extraction success and ensure enough material for PCR.

๐Ÿ“ˆ PCR amplification

โ†’ Use primers to target and amplify a barcode gene (e.g. mitochondrial 12S rRNA) via thermal cycling.

๐Ÿงซ Gel electrophoresis

โ†’ Run PCR products on an agarose gel to check for successful amplification and absence of contamination.

๐Ÿ“ฆ Sequencing

โ†’ Pool PCR amplicons, add sample-specific barcodes (indexes), and sequence using a platform like Illumina MiSeq.

๐Ÿ’ป Bioinformatics

โ†’ Demultiplex, quality-filter, and assign taxonomy to sequence data using a bioinformatics pipeline (e.g. DADA2).