EpiTect Bisulfite Kit

DNA methylation profiling Gene specific profiling - MG-63 miR-34a

Experiment
DNA methylation profiling Gene specific profiling - MG-63 miR-34a
Product
EpiTect Bisulfite Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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

Paper title
p53-Dependent Activation of microRNA-34a in Response to Etoposide-Induced DNA Damage in Osteosarcoma Cell Lines Not Impaired by Dominant Negative p53 Expression
Full paper
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Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

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