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Tissue Collection Protocol

To submit fungal tissue to our sequencing service, we require dried tissue to be placed in 1.5 mL microcentrifuge tubes. We offer tissue collection tubes in our online store. Watch the video below to learn how to collect tissue for submission.

Organize your Materials

You will need 1.5 mL tubes, tweezers, a lighter, and alcohol swabs. 

Label your Tubes

Label your tubes with the correct collection numbers. Organization is key!

Collect Tissue

Remove a piece of dried flesh the size of a grain of rice and place it in the appropriate labeled tube.  See detailed instructions below the video.

Tissue Collection Video

This video outlines the procedure to collect tissue using our preferred method in 1.5 microliter "eppi tubes."

Tissue Collection Protocol

By now you should have taken pictures of the mushroom in the field, collected the mushroom, and brought it back for preservation. Your field data slip should be filled out, providing your specimen with a unique collection number. (This is the preferred method but it is also acceptable to use the iNaturalist or Mushroom Observer observation number for labeling tubes and voucher specimens.) Your specimen should be cracker dry. We will now take a small piece of tissue from the mushroom and preserve it in a very small plastic tube: a 1.5 microliter (mL) microcentrifuge tube, also called an Eppendorf tube, or "eppi tube" for short.

NOTE: Sequencing technology and procedures continually change! Originally we used tubes with a buffer solution. Now we are putting dried tissue into empty eppi tubes. And for a while our labs wanted a small piece of specimen in a resealable bag, called a split sample, sent with the eppi tubes. We are no longer asking for splits -- although they might increase changes of success for old or degraded material. If you were sent small plastic bags along with your tubes, you can ignore them.

Lay your mushroom sample out on a clean table. Wipe off any organic matter and sterilize the tweezers with an alcohol swab and lighter. The primary goal with the flame is to burn off the alcohol, rather than getting the tweezers red-hot between each sample. A quick wipe with a swab and a very brief flame (2-3 seconds) is sufficient.

Tweeze off a small portion of mushroom tissue that is about the size of a grain of rice. It is possible to take dried tissue from any part of the mushroom in order to acquire DNA. For larger specimens, we typically suggest samples to be taken from the gills, pores or other spore-bearing surface. For small samples, we recommend taking tissue from the stem of the mushroom as to preserve as much of the cap as possible. The cap contains many more parts that could be important for future microscopic study. Thus, we want to maintain as much of it as possible intact.

Place this piece of material into your eppi tube and close the cover. Ensure that the tube is properly labeled with your collection number (field slip, iNat or MO). If you are using NAMP field slips, cut off the small number in the lower right of the slip, trim the number with scissors to remove excess border, and tape it towards the top of the label with clear scotch tape. Leave the bottom of the tube uncovered so the sample is visible. That is all that needs to be done for this sample. Insert it into the box with dividers, keeping the samples in numerical order. Repeat this process for your next sample.

Blunt tweezers are suitable for most large gilled mushrooms. For smaller specimens fine tweezers are better. For hard material like polypores and crusts a scalpel is useful. These items can usually be found at Dollar stores, arts and craft stores, or online on Amazon (for under $10).

About North American Mycoflora Project

NAMP is a 501(c)(3) non-profit organization working to create a continent-wide community of volunteer citizen scientists and professional mycologists to document the biodiversity of North American fungi.

©2017-2019 North American Mycoflora Project. All Rights Reserved.

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