I don't know if the DNA sequencing is different between dikaryotic and monokaryotic fungi. I think that is possible you will sequence the ITS region.

Hi Dr. Pertile,

thanks for the comment. Recently I sequenced a number of local edible mushroom using ITS sequence and was unable to identify a sample. I was wondering is this due to mixed species or dikaryotic fungi. Literature search didn’t give much info for me On this. Contamination was less likely because my other 10 samples results were very good.

so I am thinking maybe the mushroom sample which I mentioned just now is a mixed species or could it be a dikaryotic fungi that gives such phenomenon.

Did you make the sequencing by NGS or Sanger?

The sequencing was done by a company and our side just did the PCR and gel electrophoresis work. If not mistaken NGS was used by the company to sequence the samples.

Which primer did you use?

I was using primer that amplify specific ITS, 18s regions of Pleurotus ostreatus.

I think that the wording of your question is confusing Chong. The dna was sequenced on an abi 3730 sequencer and the its sequence does not have high background it has 2 sequences in the ITS region between the same primers that differ in over 200 bases but the pcr products are the same size

Dear Dr. Rutland,

Sorry for the confusion. I was trying to figure out will the DNA sequenced from a dikaryotic mushroom differ from a monokaryotic mushroom. The question was linked to the previous question which you had provided much enlightenment for me.

We were planning to do some DNA cloning and sequence it again. Before that is done, we are trying to figure out what are the other possibilities which can affect the sample identity, apart from mixed mushroom species, single species, hybrid or other reasons.

Regards,

Chong

Don't worry Chong it was just the use of the term background that a lot of people will assume to be random sequencing noise rather than a recognisable second sequence sequencing well under each base of the expected sequence that I was trying to clarify.These two situations will lead to different types of answer which is the situation that you would like a rationale for. Hopefully someone has some good ideas

best wishes

Paul

All right, thank you very much Dr. Paul. =)