DNeasy Blood & Tissue 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 Isolation and Bisulfite Treatment
Samples were prepared in GT buffer and stored at −20°C or −80°C. DNA was isolated using the DNeasy Blood&Tissue Kit (Qiagen, Hilden, Germany), according to the manufacturer's protocol. The tissue was homogenized by passing it through a syringe with a 20 G needle a minimum 5–10 times. The DNA was diluted in 100 μl buffer AE (Qiagen). To check quality and to exclude contamination, the DNA product was analyzed by gel electrophoresis. DNA concentrations and protein contamination of the samples were checked using a Nanodrop spectrophotometer (Isogen). Samples with a 260/230 ratio lower than 1.8 were cleaned using standard ethanol precipitation. Depending on the amount of DNA, the samples were resuspended in 20–100 μl buffer AE. After isolation, the samples were stored at 4°C until bisulfite treatment.

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Manufacturer protocol

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