BisulFlash DNA Modification Kit

DNA methylation profiling Gene specific profiling - A2780 TIMP2

Experiment
DNA methylation profiling Gene specific profiling - A2780 TIMP2
Product
BisulFlash DNA Modification Kit from Epigentek
Manufacturer
Epigentek

Protocol tips

Protocol tips
According to the predicted CpG islands and published literature, we developed one pair of nested MSP primers that is listed in Supplemental Table . Genomic DNA of ovarian cancer cells was extracted using a DNA Isolation Kit (Tiangen Biotech, Beijing, China). Bisulfite modification of DNA was performed following the recommended procedures of the BisulFlash DNA Modification Kit (Epigentek, Farmingdale, NY,USA). Placenta DNA was modified with the CpG Methyltransferase (M.SssI, NewEngland Biolabs, Ipswitch, MA, USA) and used as a positive control.

Publication protocol

Bisulfite modification and methylation-specific PCR (MSP)
To predict CpG islands in the TIMP2 promoter, we searched the TIMP2 genomic sequence including two kb upstream and one kb downstream of the transcriptional start site (TSS) using the NCBI website (). The sequences were assessed with DataBase of CpG islands and Analytical Tool (DBCAT, ) according to the following criteria: the length of the CpG island >200 bp, observed/expected CpG ratio >0.6 and percentage of G plus C >50%. According to the predicted CpG islands and published literature, we developed one pair of nested MSP primers that is listed in Supplemental Table . Genomic DNA of ovarian cancer cells was extracted using a DNA Isolation Kit (Tiangen Biotech, Beijing, China). Bisulfite modification of DNA was performed following the recommended procedures of the BisulFlash DNA Modification Kit (Epigentek, Farmingdale, NY,USA). Placenta DNA was modified with the CpG Methyltransferase (M.SssI, NewEngland Biolabs, Ipswitch, MA, USA) and used as a positive control. Every 300 ng of genomic DNA was bisulfite-treated and then quantified with a NanoDrop 2000/2000C system (Thermo Scientific, USA). Modified DNA was first amplified with flanking primers followed by specific methylated or unmethylated primers using PrimeScript™ RT Master Mix (TaKaRa, Otsu, Japan). MSP products were subjected to electrophoresis on 3% agarose gels that were stained with ethidium bromide, detected and analyzed with Image Lab Software.

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

Reviews

BisulFlash DNA Modification Kit from Epigentek 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 - A2780 TIMP2 using BisulFlash DNA Modification Kit from Epigentek.

Paper title
EZH2-mediated epigenetic silencing of TIMP2 promotes ovarian cancer migration and invasion
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Epigentek for BisulFlash DNA Modification Kit below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - A2780 TIMP2 using BisulFlash DNA Modification Kit from Epigentek. 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