MethylFlash Methylated DNA 5-mC 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

Global DNA methylation assay
The total genomic DNA was extracted from the cells (treated with GM, DM, GMAZA or DMAZA) using the Qiagen DNA Mini Kit (Qiagen Sciences, Maryland, MD) following the manufacturer’s instructions. The integrity and purity of DNA were spectrophotometrically examined according to its A260/A280 absorption. The Global DNA methylation levels were determined using MethylFlash™ Methylated DNA Quantification Kit (Epigentek, NY, USA) according to the manufacturer’s instruction. The methylated fraction of DNA is recognized by a 5-methyl-cytosine (5-mC) antibody. With this colorimetric kit, the amount of methylated DNA, which is proportional to the optical density intensity, is quantified through an enzyme-linked immunosorbent assay-like reaction (ELISA assay). The reference DNA fragments containing 5-mC and 5-cytosine are used respectively as the positive standard and negative control to generate the standard curve. In particular, manufacturer’s instruction indicates that the percentage of 5-mC in the total DNA is calculated using the following formula:Slope of the standard curve is determined using linear regression [].

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Whole genome profiling - C2C12 mouse myoblast cells using MethylFlash Methylated DNA 5-mC Quantification Kit from Epigentek.

Manufacturer protocol

Download the product protocol from Epigentek for MethylFlash Methylated DNA 5-mC Quantification Kit below.

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