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

Bisulfite DNA sequencing
Genomic DNA used for bisulfite conversion was extracted using the Wizard Genomic DNA Purification Kit (Promega, PRA1125) according to the manufacturer’s protocol. All DNA sequences for primer design were obtained from the UCSC genome browser (http://genome.ucsc.edu). Primer design and CpG island (CGI) prediction was performed using the MethPrimer software29 (http://www.urogene.org/cgi-bin/methprimer/methprimer.cgi). This software scores a DNA sequence as a CGI when it fulfills the following criteria: at least a 200-bp sequence, with a G+C content >50%, and an Obs/Exp CpG ratio >0.6. Bisulfite conversion was done using the EZ DNA Methylation Gold Kit (Zymo Research, D5005). Up to 2 μg of genomic DNA was converted using the C1000 Thermal Cycler (Bio-Rad) at 98 °C for 10 min, followed by 64 °C for 2.5 h.

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Manufacturer protocol

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

Download manufacturer protocol

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