Magattract dna blood m96 kit

Total DNA was extracted from regurgitates in a molecular diagnostic laboratory at the Institute of Ecology, University of Innsbruck, Austria. Each regurgitate sample was mixed with 200 μL lysis buffer containing 5 μL Proteinase K (10 mg/mL, AppliChem, Darmstadt, Germany) and TES-buffer (0.1 M TRIS, 10 mM EDTA, 2% SDS, pH 8) and was incubated at 56°C for 3 h. The DNA was extracted from the lysate on a BioSprint 96 robotic DNA extraction platform using the MagAttract DNA Blood M96 Kit (Qiagen, Hilden, Germany). Four negative extraction controls (DNA extraction blanks) were included to monitor for carry-over DNA contamination during the extraction process and were subsequently tested in PCR reactions for NGS.

Genomic DNA was extracted from caudal fin clips and preserved in 96% EtOH at −20°C, using a Blood & Tissue Kit (Qiagen, Hilden, Germany). From 30 μl cleared lysate, total genomic DNA was isolated using GenoM‐48 Robotic Workstation (GenoVision, Oslo, Norway) and magnetic bead technology. Binding of DNA to magnetic beads (Qiagen) was performed in 200 μl buffer MDL (MagAttract DNA Blood M96 kit; Qiagen) after which beads were washed twice in 200 μl of 80% EtOH, GenoPrep wash solution (GenoVision, Toyobo Kita‐ku, Osaka, Japan) and water, before finally eluting DNA in 0.1× TE buffer at pH 8.0. The purity and concentration of the DNA samples were determined spectrophotometrically using a NanoDrop ND‐1000 (NanoDrop Technologies, Wilmington, DE, USA).