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 Tubes containing 1x lysis buffer
20 Single use forceps
In order to avoid cross-contamination, it is important to wear clean gloves and use clean equipment for every sample.
Choosing a sample:
Red muscle tissue is preferred for consistent results but we can process organ tissue such as heart and brain. 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. Freezing at -20C or colder is preferred for long-term storage of tissue. Tissue does not need to be stored in ethanol and should not be stored in isopropanol or formalin.
Sampling Instructions (for tissue sampling kit):
•Download the Sample Submission Excel spreadsheet under the “Sample Submission” tab at www.nrdpfc.ca (you will need a password to access the download and upload functions). Access specific project spreadsheets (e.g. Wolf/Coyote Sample Submission) by clicking on the dropdown in the “Sample Submission” tab.
•Use a permanent marker to label the sample tube with a unique identifier and add this ID to the spreadsheet under Id. Then place the cursor in the “Sample Barcode” cell beside the “Id” cell and scan the barcode label on the tube. Repeat this for each new sample and complete as much of the spreadsheet as possible for each sample. The Id (your code) and the Sample Barcode (our code) will be used for cross-referencing data associated with each sample so these two labels must be linked in the spreadsheet. When the spreadsheet is complete upload the spreadsheet via the “Sample Submission” tab.
•Use clean latex or nitrile gloves to handle the sample, and with a clean scalpel blade shave off a small portion from the outside of the tissue and discard. If the sample has been stored in ethanol, minimize the transfer of ethanol to the buffer. If possible, soak the ethanol preserved tissue in Ultra-pure DNAase/RNAase-free water (e.g. Gibco, Invitrogen) or blot the tissue dry with a Kimwipe, or both.
•Then cut a piece of tissue approximately 50 mg in mass (approx. 5mm x 5mm x 2mm in dimensions) from the centre of the tissue. Note that more tissue IS NOT better because there is not enough buffer to digest a larger piece of tissue. An excessive amount of tissue will rot and DNA integrity will be compromised.
•Add the sample to the tube with preservative buffer making sure the entire piece is submerged.
•Ensure that tube lid is screwed on tight.
•Tubes can be stored and shipped at room temperature. Excess tissue sample should be kept frozen at the respective facility.