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MethylMagnet® mCpG DNA Isolation Kit
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Tools for capture and detection of methylated DNA |
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Overview
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The MethylMagnet® CpG DNA Isolation Kit is used to isolate methylated DNA from a genomic sample. The MethylMagnet® kit utilizes the MethylMagnet® GST-MBD fusion protein and glutathione magnetic beads for capture of methylated DNA. It can be used to isolate mCpG DNA from 1 ng to 1 µg of genomic DNA, or it can be scaled up to isolate much larger quantities. After capture, methylated DNA can be eluted several ways.
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How it Works
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Step 1: DNA Fragmentation
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DNA must first be fragmented so that the region of interest is physically separated from other regions on DNA that may be methylated. The island detection assays that utilize abscription™ for detection using the MethylMeter® kits have been designed to work with DNA that has been cut with the restriction endonuclease Mse I. Mse I cuts at the sequence TTAA and therefore is not affected by CpG methylation. However, MethylMagnet® can be used with DNA that has been fragmented by other methods or by restriction with other enzymes.
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Step 2: Capture of Methylated DNA
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The fragmented DNA is first incubated with the MethylMagnet® GST-MBD protein which has already been attached to glutathione magnetic beads. The DNA population will generally contain a mixture of CpG islands which are methylated and those that are not. Those which are methylated will bind preferentially to the beads via interaction of the mCpG sites and the MBD domain. Incubation is for 1 hour at room temperature. After binding, the column is washed to remove any non-specifically bound DNA.
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Step 3: Elution of Methylated DNA
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