MasterPureTM-Yeast DNA Purification kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Use prewarmed TE buffer to elute the DNA

Publication protocol

DNA Extraction
After paraffin removal using octane, DNA was extracted using the MasterPure™ Yeast DNA Purification Kit (Epicentre® Biotechnologies, Madison, WI) with an additional bead beating step to optimise fungal cell lysis as described previously [6]. This kit utilizes a non enzymatic approach to lyse fungal cell walls at 65°C. Contaminating macromolecules were then removed by a precipitation step, followed by DNA precipitation with isopropanol for purification of DNA. Extracted DNA was dissolved in 75 μl of 0,5% Triton X and stored at −20°C until PCR testing.

In order to evaluate the effect of thermal energy on the amount of amplifiable DNA, two modifications of the previously described protocol were studied. First, the initial incubation step in lysis buffer was carried out at either 65°C, as proposed by the manufacturer, or at 90°C for one hour with aliquots of all samples. Second, the duration of incubation at 90°C was increased from one hour to three, six and twelve hours using the samples 1,4,6 and 7 in duplicate extractions.

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Manufacturer protocol

Download the product protocol from Epicentre for MasterPureTM-Yeast DNA Purification kit below.

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