QIAamp DNA Blood Mini Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Publication protocol

DNA extraction.
For ocular samples, we extracted toxoplasmic DNA from up to 10 μl of the ocular sample using the QIAmp DNA Blood Minikit (Qiagen). For blood, we used two different extraction protocols. The initial protocol was performed using 200 μl of whole blood and the QIAmp DNA Blood Minikit. We followed this up with a second protocol using 2 ml of whole blood and the QIAmp DNA Blood Midi kit (Qiagen). PCR performed with extract retrieved from 200 μl of whole blood was called SV-PCR (small volume), and that performed with extract retrieved from 2 ml of blood was named LV-PCR (large volume). DNA was eluted with 100 μl of elution solution for SV-PCR and 300 μl for LV-PCR. The albumin gene was amplified as a control for extraction. After DNA extraction, samples were kept at +4°C for up to 48 h or frozen at −35°C for use within the week.

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Manufacturer protocol

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