Environmental DNA (eDNA) Sample Shipping Protocol

Clean bench area with 10% bleach, 70% ethanol, and RNase Away

Wipe down 20 µL multichannel pipette and several PCR tube holders with RNase away. Run in UV Crosslinker for 10 minutes on each side.

Wipe down ice bin with 10% bleach, 70% ethanol, and RNase Away, fill with ice, and add PCR products stored in 8-strip PCR tubes

Label all 96-well plates needed for storing samples accordingly

Being careful not to open PCR tubes directly over the 96-well plates, add 15 µL of sample to each well using multichannel pipette, making sure you keep track of the orientation of how the plate was filled

Seal the plates carefully with aluminum adhesive seals

Press around the edges of each well to make sure they are sealed

Parafilm around the edges of each plate

Wrap each plate in Kimwipes to absorb any condensation that occurs during shipping

Cut two pieces of cardboard that fit on either side of each plate; tape the cardboard around each plate to form a sandwich (to keep it from getting punctured while shipping)

Seal each plate in a new Ziploc bag and label accordingly

Bubble wrap around each plate and place in a hard container (cardboard freezer box, pipette tip box, etc.)

Seal hard container with tape and place in its own Ziploc bag; label accordingly

Place bagged box in a Styrofoam cooler that fits in an associated cardboard box and fill with ice packs

Tape Styrofoam cooler closed and secure it in the corresponding cardboard box

Store cardboard box at -20°C until ready to mail