EZ DNA Methylation-Gold Kit

DNA methylation profiling Gene specific profiling - MCF-7 BMP2

Experiment
DNA methylation profiling Gene specific profiling - MCF-7 BMP2
Product
EZ DNA Methylation-Gold Kit from Zymo Research
Manufacturer
Zymo Research

Protocol tips

Protocol tips
For best results, the CT Conversion Reagent should be prepared freshly and used immediately following preparation.

Publication protocol

Methylation-specific PCR (MSP) and bisulfite genomic sequencing
Genomic DNA of MCF-7 or MCF-7/ADR was extracted using a TIANamp Genomic DNA kit (Tiangen Biotech (Beijing) Co., Ltd., Beijing, China). Bisulfite conversion of genomic DNA was performed using the EZ DNA Methylation-Gold kit (Zymo Research, Irvine, CA, USA) according to the manufacturer’s instructions. MSP was performed on bisulfite-modified DNA. The primer sequences of BMP-2 for the unmethylated reaction were: Forward: 5′-GGGTTAGTGTTGAGTGGATTATT-3′ and reverse: 5′-CACAACACTAAAAATCAACTCC-3′ and for the methylated reaction forward: 5′-TTAGCGTCGAGTGGATTATC-3′ and reverse: 5′-GACGCTAAAAATCGACTCC-3′. PCR products were analyzed by electrophoresis in a 2.0% agarose gel containing golden view (SBS Genetech Co., Ltd., Beijing, China). For bisulfite sequencing of DNA samples, sodium bisulfite-treated genomic DNA was amplified using the following primers: Forward: 5′-GTGGTTTTTGTTGTTYGG-3′ and Reverse: 5′-TCTACCTTACTCCAATACACCC-3′. The PCR reaction products were gel purified and cloned into the pGEM-T Vector system according the manufacturer’s instructions (Promega Corporation, Madison, WI, USA). The colonies were sequenced on a genetic analyzer (Applied Biosystems, Foster City, CA, USA) to analyze the methylated cytosine level.

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Discussion

Discussion

5 years ago

Author: Jody Hancock Canada

How can I preserve leukocytes for DNA methylation profiling?

I would like to preserve leukocytes for future epigenetic analysis. How can I preserve them effectively in order to perform DNA methylation profiling at a later time?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Gene specific profiling - MCF-7 BMP2 using EZ DNA Methylation-Gold Kit from Zymo Research.

Paper title
Aberrant promoter DNA methylation inhibits bone morphogenetic protein 2 expression and contributes to drug resistance in breast cancer.
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Manufacturer protocol

Download the product protocol from Zymo Research for EZ DNA Methylation-Gold Kit below.

Download PDF Download manufacturer protocol

Videos

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