Wizard® Genomic DNA Purification Kit

Protocol tips

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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

Methyl-DNA immunoprecipitation
Genomic DNA was isolated from confluent and quiescent HUVECs using the Wizard genomic DNA isolation kit (Promega). Methyl-DNA immunoprecipitation (MeDIP) was performed with the aid of a commercial kit according to the manufacturer’s protocol (Active Motif, Carlsbad, CA, USA). In brief, 20 µg of DNA was sheared into ~ 200–400 bp fragments and immunoprecipitated with a 5-methylcytosine-specific antibody at 4 °C overnight. Immunoprecipitated DNA was bound to magnetic beads, washed and eluted. Input (sheared DNA) and immunoprecipitated DNA (1 µl) was used to amplify specific promoter regions from the gene using SYBR green PCR, as previously described. MeDIP real-time PCR primers are shown in , see section on given at the end of this article. Promoter methylation levels were estimated as immunoprecipitated DNA/total (input) DNA × 100.



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Manufacturer protocol

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

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