Before you can answer that question, tell us about the positive (and negative) controls you ran, as well as the conditions (temperature, for example - did you run a gradient?). This will allow you to draw a conclusion from the PCR reaction you performed.

you can use of PRIMER bank and primers derived from articles this is a very good option to it

Yes, prof. Lori. I run gradient based on primer temperatures. Thank you. 

We always use the following primer combination: for the ITS1 region primer pairs ITS5/ITS2, for the ITS2 region primer pairs ITS3/ITS4 described by Hinrikson et al. JCM 2005 43:2092.

Dear Ali Zuhaira,

You can go with the ITS1 and ITS4 primers as they are the Universal primers for the Fungal Identification. Here is the procedure i m writing for you for 10uL PCR reaction.

PCR Method

10x Buffer=1uL

25mM Mgcl2=0.6uL

dNTP mix= 0.2uL

Taq= 0.1 (I used Banglure GENE COMPANY)

PF(ITS1)=0.2uL (10pM Cnoc)

PR(ITS4)=0.2uL (10pM Cnoc)

MilliQ WATER=7.1uL

Cyclic Conditions

Initial Denaturation

94 Degree-5 Min

94 Degree-30 Sec

55 Degree-45 Sec

72 Degree-1 Min

35 Cycles

72 Degree-5 Min

4 Degree=Infinity (If you are going to run the PCT at Night Time)

Hope you will get the Best Results with the Mthod

Best Regards,

Srinivas

Genomic DNA=0.6uL

Hi 

Please check your DNA Extraction carefully. Aspergillus niger produces a lot of pigments which usually inhibit PCR amplification.  Use PCR enhancers and check  + and - controls  during PCR runs.

Hi , I agree with Srinivas rao Rapeti answers ...good luck.