EpiTect Bisulfite Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Always include fully methylated (in-vitro methylated DNA) and non-methylated controls (Leucocyte DNA, DNMT1/3 double KO DNA) during MSP

Protocol tips
Always include fully methylated (in-vitro methylated DNA) and non-methylated controls (Leucocyte DNA, DNMT1/3 double KO DNA) during MSP

Publication protocol

2.5 Methylation-specific polymerase chain reaction (MSP)
DNA was extracted from OS cell lines by standard method. DNA was treated with bisulfite by EpiTect Bisulfite Kit (Quiagen, Hilden, Germany) to determine aberrant miR-34a promoter methylation status. The procedure comprised different steps: bisulfite-mediated conversion of unmethylated cytosines; purification and elution of DNA and finally amplification of purified DNA by polymerase chain reaction (PCR). Primers used for methylated methylation-specific polymerase chain reaction (M-MSP) and unmethylated methylation-specific polymerase chain reaction (U-MSP) designed for the CpG area upstream of the miR-34a promoter: U-MSP 34a Reverse: 5′CAA-ACA-AAA-CAC-ATA-AAA-ACA-ACA-3′, U-MSP 34a Forward: 5′GGG-GAT-GAG-GAT-TAG-GAT-TTT-3′, M-MSP 34a Reverse: 5′ACA-AAA-CGC-ATA-AAA-ACG-ACG-3′, M-MSP 34a Forward: 5′GGG-GAT-GAG-GAT-TAG-GAT-TTC-3′. PCR products were analyzed on 2% agarose gel.



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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 - MG-63 miR-34a using EpiTect Bisulfite Kit from Qiagen.

Manufacturer protocol

Download the product protocol from Qiagen for EpiTect Bisulfite Kit below.

Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - MG-63 miR-34a using EpiTect Bisulfite Kit from Qiagen. Please note that these videos are representative and steps or experiment specific processes must be kept in mind to expect desired results.