EpiTect Bisulfite Kit

DNA methylation profiling Gene specific profiling - SiHa miR-375

Experiment
DNA methylation profiling Gene specific profiling - SiHa miR-375
Product
EpiTect Bisulfite Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

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

MSP assay
Genomic DNA was extracted from samples with a DNeasy tissue kit (Qiagen). Bisulfite reaction was performed using 5 μg of genomic DNA with the EpiTect Bisulfite Kit (Qiagen) according to manufacturer’s instruction. Then MSP was performed following the methods introduced in one previous study [] with methylated-specific primers (M): forward, 5’-TTTTCGTTTTCGTTATTAAGGTTTC-3’ and reverse, 5’-GACCCAACCTAACTTAACTTCGTC-3’ (product size: 239bp); and unmethylated-specific primers (U): forward, 5’-TTTTTGTTTTTGTTATTAAGGTTTT-3’ and reverse, 5’- CAACCCAACCTAACTTAACTTCATC-3’ (product size: 240bp). PCR products were verified by 2.5% agarose gel electrophoresis.

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Reviews

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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?

Share your thoughts or question with experts in your field by adding a discussion!

Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA methylation profiling Gene specific profiling - SiHa miR-375 using EpiTect Bisulfite Kit from Qiagen.

Paper title
MiR-375 Is Epigenetically Downregulated by HPV-16 E6 Mediated DNMT1 Upregulation and Modulates EMT of Cervical Cancer Cells by Suppressing lncRNA MALAT1
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

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - SiHa miR-375 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.

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