Does anyone have any knowledge of PCR inhibitions by Prepman extraction vs MagNA Pure Extraction?

Marc-Antoine Perrenoud @Marc-Antoine-Perrenoud

20 February 2015 1 565 Report

I'm currently running PCR for fungal (18S gene, 26S gene). My PCR for 18S and 26S works fine on fungi (extracted with MagNA Pure 96 from Roche), but I can't get amplification for yeast (Cryptococcus neoformans, Candida lusitaniae), extracted with Prepman protocol.

My suspicion is that something in my Prepman Extraction is inhibiting my PCR but I could be wrong.

Does anyone have experience with fungal PCR and could help me solve this problem?

Cheers

Ziguo Zhang

It was really a very long time ago when I met such problem. I remember I included some non-fat milk in the reaction to overcome the inhibition. Peg precipitation your template DNA also helps.

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