I think, ITS 86F/ITS 4 primer pair is the best for studying fungal diversity as well as discrimination of fungal species.

J. C. Tarafdar Thank you kindly for info.

The primers best work for the said purpose are ITS1 forward

(5'-TCCGTAGGTGAACCTGCGG-3') and ITS2 reverse (5'TCCTCCGCTTATTGATATGC-3'). They usually amplify ~450-550 bp fragment. It will surely resolve the identity of organism till genus level. However, for species level identification you should try to amplify D1/D2 variable region. For primer sequences and details of PCR condition please refer:

https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0159043

Dear Huseyin

Application of PCR in mycology Edited by Bridge ,P.D et at . may be usuful for you.

Best Regard

Ejaj K Pathan and Mohammed Fayyadh thank you kindly for your advice.

I found that the primers ITS1F/ ITS4 and LROR/ LR7 are the best for fungal indentification. They are so usefull.

I used the universal primers ITS1/ITS4 with some difficulties.

Good luck.

Sana Hamrouni Thank you kindly for advice.

ITS1 and ITS2 are the regions that are usually targeted for Illumina sequencing.

In my experience, I have always used the primers ITS1F/ITS2R targeting the ITS1 sublocus.

However, it depends on what are your questions and your start material (soil, roots, water,...)

For soil and plant material have a look a this publication

Article Drivers of total and pathogenic soil-borne fungal communitie...

With ITS2 you can have plant co-amplification (it depends on the plant species), but you can detect the most abundant members of the Glomeromycota in your samples. The ITS1 primers usually have a low resolution for the AMF.

Davide Francioli thank you for kindly advice.

Dear, Hüseyin Burak Dişli,,

There are many primers depending on your sustances or materials. So the best primers are ITS1, ITS2, ITS4, etc...

Regards