EZ DNA Methylation 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.

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

Publication protocol

Methylation-specific polymerase chain reaction
For methylation analysis, 2 μg of genomic DNA obtained from gynecologic cancer cell lines were modified using the EZ DNA Methylation™ Kit (Zymo Research, Orange, CA, USA). Primers specific for bisulfite modified DNA were designed using MethPrimer software (http://www.urogene.org/methprimer/index1.html). The primers used in this study (located −8061 to −7782 from transcription start site, modified from Pleshkan et al. [12]) were as follows: CD133 M primer sense (5′-TTCGGGATAGAGGA AGTCGTAA-3′) and CD133 M primer antisense (5′-CTCCCGCCCTAATCACCGCT-3′); and CD133 U primer sense (5′-TTTGGGATAGAGGAAGTTGTAA-3′) and CD133 U primer antisense (5'-CTCCCACCCTAATCACCACT-3') (Figure ​(Figure1).1). The PCR conditions were as follows: 94°C for 5 minutes, followed by 43 cycles of 94°C for 30 seconds, 60°C (methylation-specific PCR, MSP) or 62°C (unmethylation-specific PCR, USP) for 30 seconds, and 72°C for 60 seconds, and finally 72°C for 7 minutes. The amplified DNA fragments were fractionated on 2% agarose gels and stained with ethidium bromide.

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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 - OVCAR-3 human ovarian cancer using EZ DNA Methylation kit from Zymo Research.

Manufacturer protocol

Download the product protocol from Zymo Research for EZ DNA Methylation kit below.

Download manufacturer protocol

Videos

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