Genomic DNA extraction
Genomic DNA was extracted from samples containing a pool of fish eggs and larvae fixed in ethanol. To ensure the complete evaporation of the alcohol, initially the excess was removed through pipetting, and then the microtubes were kept open for three hours at 55°C. We then added 600µl of TNES buffer to each sample and ground the bulk with a plastic pestle until only minuscule tissue fragments were left. Next, 20µl of proteinase K (20mg/ml) was added to each microtube. The samples were kept at 55°C until complete tissue digestion. Finally, the genomic DNA was extracted using a low-cost saline protocol adapted from (Aljanabi and Martinez, 1997).
The samples were quantified using a Qubit 4 Fluorometer (Thermo Fisher) with a 1x dsDNA HS Assay Kit (Thermo Fisher) to verify the success of the DNA extraction. The samples were then diluted to 100ng/µl.