Description
This kit contains the materials needed to uniquely identify fungal species using DNA technology. There are hundreds of thousands of known species of wild mushrooms and other fungi in the world. And millions more that we are yet to discover! Fungi can be found in virtually all known ecosystems, yet many species look similar to each other, making them difficult to identify by eye.
Start with your own samples, from the mold on your overripe tomato to wild-harvested fruiting mushrooms. Once you have extracted their DNA, you will be able to amplify 80 unique fungal DNA barcodes by PCR and, with the help of an add-on kit and a sequencing service, obtain DNA sequences for up to 40 of them.
- Techniques: Micropipetting, DNA extraction, PCR, gel electrophoresis, bioinformatics
- Topics: Biodiversity monitoring, DNA barcoding, species identification
- Time required: This is a project-based activity that requires at least three class periods for completion
- Level: Advanced high school, college, independent research
This is an advanced lab activity with unknown outcomes and is ideal for independent research projects. Thumbnail photo by Volodymyr Tokar.
This lab kit provides enough reagents for 80 PCR experiments:
- 2X PCR Master Mix, Clear
- ITS1/ITS4 Primer Mix
- DPX™ Buffer for DNA extraction
- Fast DNA Ladder 1
Supplied by user:
Standard reagents and consumables must be supplied by the user. The Learning Lab Companion Kit (catalog no. KT-1510-01) contains all the plasticware and gel electrophoresis reagents needed, or these can also be sourced independently.
Reagents and consumables supplied by user:
- Agarose (electrophoresis grade)
- DNA stain (e.g., SeeGreen™ or GelGreen®)
- Gel electrophoresis buffer (e.g., 1X TBE)
- Distilled or deionized H2O (to dilute buffer concentrate – not included in Lab Companion Kit)
- Microtubes
- PCR tubes
- Fungal samples
Required equipment:
- Micropipettes and tips
- Tissue homogeneizer or vortex mixer with microtube holder attachment
- Microcentrifuge
- Thermal cycler
- Gel electrophoresis and visualization system
- Sanger DNA sequencing services
Storage notes:
- Reagents may ship at room temperature, but require freezer storage upon arrival
- Frozen reagents are stable for 12 months when stored in freezer
Fungal Sequencing Add-On
This kit is an add-on to the Fungal DNA Barcoding Kit (MP-KT-1014-01). It contains primers, nuclease-free water, and tubes aimed to facilitate submission of amplified ITS PCR products for Sanger sequencing.
Contents:
- ITS1 Primer, 5 µM (200 µL)
- ITS4 Primer, 5 µM (200 µL)
- Nuclease-Free Water
- 12 strips of 8 PCR tubes and caps
Please follow specific service guidelines for sample submission. Note that some services will also supply ITS1 and ITS4 primers for a fee.
Storage notes:
- Reagents may ship at room temperature, but require freezer storage upon arrival
- Frozen reagents are stable for 12 months when stored in freezer
