Wizard® Genomic DNA Purification Kit

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	For blood DNA isolation, blood must be collected in EDTA, heparin or citrate anticoagulant tubes to prevent clotting.

Protocol tips
For blood DNA isolation, blood must be collected in EDTA, heparin or citrate anticoagulant tubes to prevent clotting.

Publication protocol

Wizard genomic DNA purification kit with
modifications The wizard genomic DNA purification kit (Promega/USA) with several modifications was used as follows: For all DNA preparations, cells were grown in the corresponding medium containing 1.5% glycine to facilitate cell lysis. An overnight culture of 5 ml was pelleted by centrifugation at 14,000 rpm for 3 min at 25°C and re-suspended in 900 µl of 50 mM EDTA. A volume of 120 µl of solution of lysozyme [(40 mg/ml) Sigma Chemical Co.USA] was added to the cell suspension and incubated in a water bath for 1 h at 37°C with occasional mixing. The suspension was centrifuged (14,000 rpm at 25°C for 3 min) and the pellet was gently re-suspended in 900 µl nuclei lysis solution (promega). To induce lysis, the cells were incubated at 80°C for 5 min. After cooling at room
temperature, 4 µl of RNase A (50 mg/ml, Sigma/USA)
were added and the tubes were inverted 10 times before the mixture was incubated again for 1 h at 37°C with occasional inversion. For protein precipitation, 300 µl precipitation solution (promega) was added to the mixture and vortexed vigorously for 30 s, and then, the mixture was incubated in ice for 7 min and centrifuged at 14,000 rpm for 15 min). The supernatant was carefully transferred to clean 1.5 ml Eppendorf tube containing 600 µl isopropanol at room temperature and the mixture was gently mixed by inverting the tube followed by centrifugation at 14,000 rpm for 10 min. The pellet was washed with 70% ethanol to remove residual contaminants. Finally, the ethanol was discarded and the pellet containing the genomic DNA was re-suspended in 60 µl DNA rehydration solution (Promega).

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Manufacturer protocol

Download the product protocol from Promega for Wizard® Genomic DNA Purification Kit below.

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