eDNA extraction
@Zerina Rahic
Extraction process begins when multiple samples are captured via filtration or precipitation
- May occur in the field or laboratory facilities (cold storage or ambient storage with ethanol)
- Filtration is preferred for high sample volumes (use of ethanol precipitates means less water can be processed at one time)
- Homogenization of filtration paper via ceramic bead tubes to facilitate optimal sample lysis
- Homogeneous solution is then suspended in a transfer lysate
- eDNA is extracted by highly specific binding of nucleic acids using superparamagnetic beads
- Extract is washed and eluted to yield a high purity eDNA extract to be used for the desired downstream application
- Downstream application:
- Polymerase chain reactions (PCR), including end-point, quantitative, and digital droplet PCR.
- High-throughput sequencing (HTS) methods, and subsequent bioinformatic analyses.
Two figures below summarize eDNA extraction
References