A. Sampling Section (Protocol optimized for freshly collected mussels)
Be sure to gather all materials before collecting mussels.
B. Extraction of circulating cell-free DNA Section
1) Equilibrate samples to room temperature (15–25°C).
2) Equilibrate Buffer ATE to room temperature.
3) Set a thermomixer or heated orbital incubator to 56°C for use in step #13.
4) Ensure that buffers AW1 and AW2 have been prepared :
a) Add 25 mL ethanol (100%) to the bottle containing 19 mL Buffer AW1 concentrate.
b) Add 30 mL ethanol (100%) to the bottle containing 13 mL Buffer AW2 concentrate.
5) If Buffer AL or Buffer ATL contains precipitates, dissolve by heating to 70°C with gentle agitation.
6) Reconstitute carrier RNA by adding 310 µL of buffer ATE.