EpiTect 96 Bisulfite Kit (2)

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	Follow manufacturer's instructions

Protocol tips
Follow manufacturer's instructions

Publication protocol

For pooled dried blood samples, sodium bisulfite treatment was performed with the EpiTect 96 Bisulfite Kit (QIAGEN) in accordance with the manufacturer's protocol. This procedure was found to result in a higher yield of DNA after bisulfite treatment than the traditional homebrew method described above. In brief, 20 μl of extracted DNA was treated with 85 μl of bisulfite solution and 35 μl of DNA protect buffer. The sample was incubated on a thermal cycler under the following conditions: initial denaturation at 99°C for 5 min, followed by 60°C for 25 min, 99°C for 5 min, 60°C for 85 min, 99°C for 5 min, and 60°C for 175 min. After sodium bisulfite treatment, 560 μl of BL solution with 10 mg/ml of carrier RNA was added, and the sample was transferred to the 96-well binding plate and pulled through the columns with a vacuum. The columns were washed with 500 μl Wash Buffer. The bisulfite-treated DNA was desulfonated on the column by the addition of 250 μl of BD solution and incubation at room temperature for 15 min. The desulfonated DNA bound to the column was washed two times with Wash Buffer followed by one wash with 95% ethanol. The columns were dried for 10 min under the vacuum. The 96-well binding plate was transferred to a collection plate, and 70 μl of Buffer EB and 10 μl of Top Elute were added to each column. A vacuum was applied for 1 min for elution of the DNA. The bisulfite-treated DNA was stored at −80°C.



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Manufacturer protocol

Download the product protocol from Qiagen for EpiTect 96 Bisulfite Kit (2) below.

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