EpiTect MSP Kit

DNA methylation profiling Gene specific profiling - Ca Ski MEG3

Experiment
DNA methylation profiling Gene specific profiling - Ca Ski MEG3
Product
EpiTect MSP 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

Methylation specific PCR for promoter of MEG3
First, DNA from cells and tissues was extracted using the QIAmp DNA Mini kit (Qiagen, German), according to the manufacturer’s recommendations. Next, we used the EpiTect Bisulfite kit (Qiagen) for bisulfate conversion following the product manual. At last, Methylation specific PCR (MSP) was performed on Bisulfite-treated DNA using EpiTect MSP kit (Qiagen). The primers of MEG3 promoter which specific for methylated and unmethylated were as follows: the methylated pair (M) was 5′-GTT AGT AAT CGG GTT TGT CGG C (forward) and 5′-AAT CAT AAC TCC GAA CAC CCG CG (reverse); the unmethylated pair (U) was 5′-GAG GAT GGT TAG TTA TTG GGG T (forward) and 5′-CCA CCA TAA CCA ACA CCC TAT AAT CAC A (reverse). The PCR reaction was conducted in DNA Engine Tetrad 2 Peltier Thermal Cycler (Bio-Rad) under the following conditions: 95 °C 15 min; 94 °C 30s, 70 °C 30s, 72 °C 30s, 5 cycles; 94 °C 30s, 65 °C 30s, 72 °C 30s, 5 cycles; 94 °C 30s, 60 °C 30s, 72 °C 30s, 30 cycles; 72 °C 7 min. The PCR products were identified by electrophoresis through 2.5% agarose gel, stained with ethidium bromide, visualized under UV and compared by densitometry.

Full paper   Login or join for free to view the full paper.

Reviews

EpiTect MSP Kit from Qiagen has not yet been reviewed for this experiment

We'd love it if you would be the first to write a review!

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 - Ca Ski MEG3 using EpiTect MSP Kit from Qiagen.

Paper title
Downregulation of long noncoding RNA MEG3 is associated with poor prognosis and promoter hypermethylation in cervical cancer
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

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

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - Ca Ski MEG3 using EpiTect MSP 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.

We haven't found any additional videos for this experiment / product combination yet.

Outsource your experiment

Fill out your contact details and receive price quotes in your Inbox

  Outsource experiment
Become shareholder Discussions About us Contact Privacy Terms