QIAamp DNA 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

T7 Endonuclease 1 (T7E1) assay
After TALEN transfection, cells were split and partly used for genomic DNA extraction using the QIAamp DNA Blood Mini Kit (Qiagen) following the manufacturer's instructions. A PCR amplifying the spacer locus was carried out using the primers shown in Table 3. For one reaction, 10 μl 5× HF buffer, 1.5 μl 50 mM MgCl2, 1 μl 25 mM dNTP mix, 2 μl 10 mM primer each, 0.5 μl Phusion High‐Fidelity DNA Polymerase (NEB) and 100 ng genomic DNA were used, filled up to 50 μl with HPLC‐grade H2O. PCR was performed under following conditions: initial hotstart denaturation at 98°C for 5 min, followed by 35 cycles with 98°C for 30 s, 60°C for 30 s, 72°C for 30 s and finished by a final extension step at 72°C for 5 min in a PCR cycler (Peqlab). PCR products were analysed by gel electrophoresis and purified using QIAquick PCR purification kit (Qiagen).

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Manufacturer protocol

Download the product protocol from Qiagen for QIAamp DNA Mini Kit below.

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