EZ DNA Methylation-Direct Kit

DNA methylation profiling Gene specific profiling - A2780 miR-30c-5p

Experiment
DNA methylation profiling Gene specific profiling - A2780 miR-30c-5p
Product
EZ DNA Methylation-Direct Kit from Zymo Research
Manufacturer
Zymo Research

Protocol tips

Upstream 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 (MSP)
DNA bisulfite modification and purification were performed using an EZ DNA methylation-Direct kit (Zymo Research Corporation, Irvine, California, USA) according to the instruction. Concentration of DNA was evaluated by absorbance at 260 nm on a UV spectrophotometer (BioRad Inc., Hercules, CA, USA). The sets of primers for miR-30a/c-5p gene were flanking the 3kb regions of the 5’ upstream regions from the start of pre-miR-30a/c-5p sequences. The primers for methylation-specific PCR were designed by MethPrimer and the sequences were as follows: miR-30a methylated (M)-forward (F): 5’-TAAATATTTTAAGGATAGAGAGCGA-3’, miR-30a methylated (M)-reverse (R): 5’-AAAATTTAAAAACAACCCGAA-3’, miR-30a unmethylated (U)- forward (F): 5’-AAATATTTTAAGGATAGAGAGTGA-3’, miR-30a unmethylated (U)- reverse (R): 5’-ATAAAAAATTTAAAAACAACCCAAA-3’; miR-30c methylated (M)-forward (F): 5’-AGGCGTTTGTAGTTTTAGTTATTCG-3’, miR-30c methylated (M)-reverse (R): 5’-CGAAATAAAATCTCGCTCTATCG-3’, miR-30c unmethylated (U)- forward (F): 5’-GTGTTTGTAGTTTTAGTTATTTGG-3’, miR-30c unmethylated (U)- reverse (R): 5’-TATACTCCAACTACTCACACCACG-3. DNA amplification was performed with Epi Taq HS (Takara Biotechnology Co. Ltd., Dalian, China) under the following condition: 94 °C for 5 min; 40 cycles of 94 °C for 45 s, 44.3 °C (miR-30a U)/48 °C (miR-30a M)/36 °C (miR-30c U)/50 °C (miR-30c M)/for 45 s, 72 °C for 1 min; and 72 °C for 10 min. The PCR products were separated by 2.0 % agarose gel electrophoresis and visualized by a chemiluminescence imaging system (Bio-Rad, Richmond, CA, USA).

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

Reviews

EZ DNA Methylation-Direct Kit from Zymo Research 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 miR-30c-5p using EZ DNA Methylation-Direct Kit from Zymo Research.

Paper title
A Feedback Loop Between miR-30a/c-5p and DNMT1 Mediates Cisplatin Resistance in Ovarian Cancer Cells.
Full paper
Login or join for free to view the full paper.

Manufacturer protocol

Download the product protocol from Zymo Research for EZ DNA Methylation-Direct Kit below.

Download PDF Download manufacturer protocol

Videos

Check out videos that might be relevant for performing DNA methylation profiling Gene specific profiling - A2780 miR-30c-5p using EZ DNA Methylation-Direct Kit from Zymo Research. 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