EZ DNA Methylation-Gold Kit

Protocol tips

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

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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Reviews

Discussion

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.

Manufacturer protocol

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

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - MCF-7 BMP2 using EZ DNA Methylation-Gold Kit from Zymo Research. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.