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    Environmental DNA (eDNA) Sample Shipping Protocol

    Alexandria B Boehm, Meghan M. Shea

    Published: 2023-08-02 DOI: 10.17504/protocols.io.dm6gp38jpvzp/v1
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    Abstract

    This is a protocol for mailing PCR amplicons generated from eDNA samples, specifically via the following earlier sampling & filtration, extraction, and PCR protocols, e.g.:

    Coastal Environmental DNA Sampling & Gravity Filtration Protocol

    DNA Extraction Protocol from Sterivex Filters

    Environmental DNA (eDNA) COI PCR Amplification and Gel Electrophoresis ProtocolWhile this protocol was designed for mailing to the Georgia Genomics and Bioinformatics Core, the general principles may be relevant to other shipping contexts.

    Acknowledgements & Attributions:

    Thanks to Bradley B. Tolar for guidance on successful shipping protocols.

    Steps

    Preparing Samples for Shipping

    1.

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

    Note
    Ideally, this should be a designated post-PCR bench area.

    2.

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

    3.

    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

    4.

    Label all 96-well plates needed for storing samples accordingly

    5.

    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

    6.

    Seal the plates carefully with aluminum adhesive seals

    6.1.

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

    7.

    Parafilm around the edges of each plate

    8.

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

    9.

    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)

    10.

    Seal each plate in a new Ziploc bag and label accordingly

    11.

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

    12.

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

    13.

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

    14.

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

    Note
    If you need to include any order information or documents alongside the samples, don't forget to put them in the box here.

    15.

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

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