Sample Collection of Tissue for DNA Analysis

Preparation of Tissue Samples for DNA Analysis:
DNA is a relatively stable molecule but will degrade over time unless preserved properly. The tissue sampling kit (NRDPFC-001) can be used to sample tissue from carcasses or animal parts. Included in the kit are the following items:

• 20 x Cryovial,
  
• 1ml round bottom filled with 500ul of 1x lysis buffer (two vials for each sample submitted),
  
• 10 Scalpels,
  
• 1 x Needle waste bottle.

In order to avoid cross-contamination, it is important to wear clean gloves and use clean equipment for every sample.

Choosing a sample:
The best samples are from the brain, heart, and red muscle. DNA in these samples will degrade in 2-3 weeks in warmer temperature. Burrowing into the tissue may be necessary to obtain the freshest sample. Kidney, spleen, liver and stomach can be collected but these tissues degrade faster than the ones listed above.

Sampling Instructions (for tissue sampling kit):
1. Label two tubes for each sample using a permanent marker
2. Using clean latex gloves to handle the sample cut a piece of tissue approximately the size of a 1 ½ cm long spaghetti noodle from the inside area of the tissue. (You want to avoid the outside portion of the tissue if at all possible as it may contain contamination and may be degraded.)
3. Add the sample into the first tube with the preservative buffer. (This is an adequate amount of sample, if you add too much it will begin to rot.)(Figure 1)
4. Repeat the same procedure a second time with the same sample for the second sample tube provided.
5. Tubes can be shipped at room temperature. Any excess tissue should be kept frozen at the respective facility.

For those people who have purchased sampling kits from the Wildlife Forensic DNA Laboratory in the past there is another possible method for sampling meat, organs and hide.

1. Label two tubes for each sample using a permanent marker.
2. Attach the 16 gauge needle to the syringe.
3. Using clean latex gloves to handle the sample, pull back on the syringe plunger until it is at the 3mL graduation.
4. Pierce the sample fully and forcefully with the needle five times.
5. Add the sample into the tube with the preservative buffer, gently as the plunger may be under pressure, by depressing the plunger with the needle facing the inner edge of the tube.
6. Repeat the same procedure a second time with the same sample for the second sample tube provided.
7. If this procedure does not provide sufficient results refer to the previous directions and use a scalpel.

Natural Resources DNA Profiling and Forensic Centre DNA Building, Trent University, 2140 East Bank Drive, Peterborough, Ontario, Canada, K9J 7B8 Phone: (705) 748-1011 ext. 7126| Fax (705) 748-1132 Email: info@nrdpfc.ca