eDNA extraction from water samples filtered through Sterivex filter units (NucleoMag DNA/RNA Water Kit - MACHEREY NAGEL). v1

The objective of this protocol is the environmental DNA (eDNA) extraction from water samples filtered through Sterivex filter units. The Sterivex units can be prefilled with preservation buffer or not. DNA extraction is performed using a MagnetaPure 32 Nucleic Acid Purification System (Dutscher) and with the NucleoMagDNA/RNA Water Kit (Macherey Nagel). The procedure is based on reversible adsorption of nucleic acids to paramagnetic beads under appropriate buffer conditions. The benefits of using the MagnetaPure 32 Nucleic Acid Purification System (Dutscher) are increased productivity and repeatability, as well as eliminating human error and the pain of repetitive work. This protocol is used prior to molecular biology analysis (e.g. qPCR, metabarcoding, ddPCR) to specifically target both macro- and micro-organisms eDNA extracted from water samples. This protocol is optimised for rare eDNA and the suggested elution volumes are therefore low (between 50 and 65 µL), but can be increased if targets are more abundant.