MethylFlash Hydroxymethylated DNA Quantification Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Ensure DNA is mixed thoroughly prior to application. The wells of the assay strip bind up to 200 ng of total DNA.

Protocol tips
Ensure DNA is mixed thoroughly prior to application. The wells of the assay strip bind up to 200 ng of total DNA.

Publication protocol

Quantification of 5mC, 5hmC
The ELISA-based ‘MethylFlash Methylated DNA Quantification Kit (Colorimetric)’ and the ‘MethylFlash Hydroxymethylated DNA Quantification Kit (Colorimetric)’ (all from Epigentek, distributed by BioCat GmbH, Heidelberg, Germany) were used to quantify the amount of 5mC and 5hmC in the DNA of GCC cell lines. Quantification was performed in triplicates according to the manual. 100 ng of total DNA was used for 5mC quantification and 200 ng for 5hmC quantification. The initial incubation time was 90 min and the final developing time was 10 min. For absolute quantifications standard curves were generated by plotting the concentration of the positive control supplied with the assay against the optical density (OD) at 450 nm after performing the assay. For relative quantification the OD at 450 nm of 5 ng/µl of assay positive control was used as reference.

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Manufacturer protocol

Download the product protocol from Epigentek for MethylFlash Hydroxymethylated DNA Quantification Kit below.

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