Dear Bhawana,

We also use the same kit in our lab. to extract DNA from pure cultures ( on PDA) for various fungi. I have not worked with Diaporthe fungus but with many others like: Colletotrichum, Fusarium, Penicillium, Septoria, Cryphonectria, Pleurotus, Tuber etc...generally I use to extract the DNA from many pure fungal cultures and then identify the fungus by PCR and ITS seqyencing and do other further studies.

So, in my experience the kit from Qiagen DNeasy mini kit is not always the best. I mean you may get sometimes very less concentration of DNA unless you make some small modifications to the protocol for example you take more starting material at the begining and adjust the buffers accordingly all over the workflow. Anyway, I can say that the final yield you get is very much species dependent ....

For some fungi you have nice yield but for others as you very less...

At present I am using another kit which I found it better for fungi extraction it gives me more than 100ng/micro liter of DNA...

You can try to elute with your kit in less volume ( around 30 micro liters sterile water of EB ) but again try to change the protocol or buy this kit:

DNA, RNA, and protein purification called NucleoSpin Plant II from Macherey-Nagel -Germany. This kit is quite universal so it can be used for plants, fungi, soil, compost , drug and animal excrements and works pretty well.

I am using it now for a while and for both fungi and compst and I can assure youn that it worked good for me.

I changed to it because for some fungi the Qiagen one you said did not give me for some fungi great yiedls even if I modified it as much as I could...

Any other help just let me know and I'll try to answer and see if you improve the yield..

Good Luck,

Stefania

Thank you Stefania.. :) I will get back to you .

I tried a new protocol by instagene matrix.

It yielded upto 150 ng/micro liter.

I hope it works.