Description
We are now offering ready-to-use, pre-made, dual-index, PCR-ready primer plates for ITS fungal barcoding. If you or your lab has a thermocycler that accepts fully skirted plates (primarily Eppendorf and Bio-Rad thermocyclers), then these plates will expedite your local barcoding efforts. All you need to do is to perform PCR locally, pool (combine) the samples, and mail in the pool. We will then sequence the sample and provide you the final results in a MycoMap project. All for about $1.00 per sample.Process:
1. Perform a DNA extraction on your fungal samples. We typically use a basic X-amp extraction, but a more detailed protocol will improve your success rates.
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. Add 0.5 uL of the DNA extract to each well of the PCR plate directly through the foil seal.
5. Fill in the spreadsheet for each sample.
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 cell into a 1.5 uL microcentrifuge tube.
9. Mail in the 1.5 uL microcentrifuge tube.
10. We perform the DNA sequencing and analysis on the final samples.
Materials Needed:
DNA extract (can be as simple as X-amp and a heat block)
10uL pipette
10uL pipette tips
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 1.5mL eppi tube
Metadata spreadsheet
Shipping to you
***Success rates will be dependent upon the PCR extraction methodology that you select. A basic X-amp 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. At this time we are not shipping internationally.