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Tissue Collection for Sequencing 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. Paid orders can order tubes from our store.

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 voucher data card should be completely filled out and your sample should have a collection number. We will now be taking a small piece of tissue from the mushroom and preserving it in a very small plastic tube: a 1.5 microliter (mL) microcentrifuge tube, also called an Eppendorf tube, or "eppi tube" for short.

Lay your mushroom sample out on a clean table. Sterilize 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.

We will begin by creating a "split" of each sample. Along with your tubes, you were sent small plastic bags for each sample. You will be placing a small portion of your collection in this bag. First, label the bag with your mycoflora number (or other collection number). If there are multiple specimens in the collection, place one of the smaller specimens in this bag. If there is only a single mushroom, place a small portion of the specimen, containing gills or pores, into the bag. The piece does not need to be larger than a quarter. For very small samples, where the total collection would be signficantly reduced or damaged by creating a split, you may skip the process of creating a split of the collection, and retain the entire collection for shipment to your herbarium. 

Tweeze off a small portion of the mushroom tissue from your split that is about the size of a grain of rice. Place this piece of material into your eppi tube and close the cover. Ensure that the tube is properly labeled with the collection number. Utilizing the small number in the lower right of your field data slip, trim the number with scissors to remove excess border, and tape it towards the top of the label with clear scotch tape. Try to leave the bottom of the tube uncovered so the sample is visible. That is all that needs to be done for this sample. Now you can "sterilize" your scalpel and tweezers (as described above) to repeat this process for your next sample.

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. 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.  

NOTE: Projects that are utilizing Duke University in North Carolina as their primary herbarium can send the specimens in-tact, without creating the split.

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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