Marwan Al-Maqtoofi First, you must confirm that your DNA can be amplified in PCR with some other primers, second, that your Β-tubulin2 primers are working on control DNA samples. After this you can look for other primer set.

Firstly, can you try ITS region which is predominantly used for molecular identification of most genus of fungi. After the genus has been identified there are other several primers for determining the species for different genera.

Hi Alex and Joshua,

I did ITS and TEF all were working perfectly. I send the plate of this fungus isolate to Macrogen in South Korea. They did gDNA extraction, ITS and TEF. All were good except B-tubulin-2 has not been amplified.

This is exact case. Is there any helpful suggestion please? ITS and TEF indicated to a new novel fungal species.

After PCR, was there amplification for the PCR products for beta tubulin region?

No

That is why I'm so confusing. Macrogen sent me the gradient PCR report with no PCR amplification

Dear Jong-hwan Shin,

I've tried RPB2 but also could not get any amplification. That is why I'm totally confused