Low fungal DNA concentration from leaf.

13 February 2013 1 2K Report

I have been working on quantifying genomic DNA extracted from leaf (sugar beet) which was inoculated with fungus. I am able to amplify sugar beet DNA while I am not able to amplify fungal DNA. The DNA from leaves were extracted 3, 5 and 7 days post inoculation. Now the strange part: I am able to amplify fungal DNA with a regular PCR but am unable to amplify fungal DNA in a qPCR (Taqman Probe). I would appreciate any comments, suggestions or any different protocol that I can follow.

I use CTAB method for DNA extraction and FAM and HEX probes for qPCR.

Keshav Birla

Thanks for your reply Touseef. I have already followed the suggestions you gave me. I did a gPCR to check the right annealing temperature and my primers for qPCR give a product less than 150 bp.

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