Figure 3: Experimental workflow. | Scientific Reports

Figure 3: Experimental workflow.

From: In silico assessment of primers for eDNA studies using PrimerTree and application to characterize the biodiversity surrounding the Cuyahoga River

Figure 3

We first isolated DNA from 40 ml of river water. We then amplified each sample with 12 taxon-specific primer sets and pooled a portion of each PCR for each sample. Each primer had a 5′ tail to allow a second PCR which added Illumina adapter sequence and an individual index. We then pooled all barcoded samples and sequenced the library on a MiSeq. We used the sequence information to identify species of origin for DNA fragments isolated from the original samples.

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