Description
Streamline your fungal barcoding efforts with our ready-to-use, pre-made, dual-index PCR primer plates.
If your lab has a thermocycler that accepts fully skirted plates (e.g., most Eppendorf and Bio-Rad models), these plates will significantly expedite your workflow. Simply perform the PCR locally, pool your samples into the provided tube, and mail it back to us. We handle the sequencing and provide final results via a MycoMap project—all for roughly $1.00 per sample.
Process:
1. Perform a DNA extraction on your fungal samples. We typically use a basic XNA DNA extraction, which gives great success rates for most groups of macrofungi, but a more detailed protocol will improve your success rates for more difficult specimens.
2. Spin down the PCR plates.
3. Wipe down the foil surface of the PCR plate with an alcohol swab and let it dry.
4. Fill in the spreadsheet for each sample.
5. Add 0.5 uL of the DNA extract to each well of the PCR plate directly through the foil seal.
6. Put the clear PCR sealing film on the plate.
7. PCR the plate on a thermocycler.
8. Add 2uL of PCR product from each of the 96 wells into a 2.0 mL microcentrifuge tube. (It is often easiest to multichannel into an 8-strip then combine.)
9. Mail in the 2.0 mL microcentrifuge tube.
10. We perform the DNA sequencing and analysis on the final samples.
Materials Needed:
DNA extract (can be as simple as XNA in a well and a heat block for 15 minutes)
10uL pipette (not included)
10uL pipette tips (not included)
Thermocycler that accepts fully skirted plates
Plate centrifuge or a salad spinner
Included in this kit:
One 96-well dual-indexed, PCR-ready primer plate with ITS1F and ITS4 primers
One PCR sealing film
One 2.0mL screw-top microcentrifuge tube
Metadata spreadsheet
Shipping to you
***Success rates will be dependent upon the PCR extraction methodology that you select. A basic XNA extraction will typically yield 70-80% success on the initial PCR attempt. A more detailed kit extraction will approach 90-100% success rates for recently collected specimens.***
We are able to offer premade plates that contain alternative fungal primers or primers for other organismal groups. Additional upfront charges may apply.
We will attempt to ship internationally, but you must be aware of your import regulations/restrictions and additional shipping charges will apply.