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Epigenetics

MDQ1 Troubleshooting Questions

1. Can I dilute my DNA with water instead of the DNA Binding Solution supplied in the Kit?
Yes. You can use water.

2. Can I dilute all of the antibody at once and save it for later?
No.  A fresh dilution of the antibody should be prepared each time the kit is used.  Diluting the antibody all at once will decrease it's stability and it's effectiveness in the kit.

3. Can you measure promotor methylation by this kit?
This kit is not suitable for measuring promotor or locus specific methylation.  It measures Global methylation.

4. What is the sensitivity of the kit?  What difference in methylation status can be detected?
Our past results suggest a 20% difference in methylation can routinely be seen.  The best answer is that one should perform replicate experiments, determine the error of measurement (technical and/or biological) and apply a p-test or other statistical method to determine how it works in your hands.

5. Are there any species restrictions for this kit?
The antibody works on me-C, so any species with DNA that methylates at 5-me-C will work.

6. Any suggestion on how to isolate the methylated DNA? Any recommendations for this step?
We use Sigma’s GenElute™ Mammalian Genomic DNA Miniprep Kit (G1N10).

7. Any suggestion on how to store methylated DNA before analysis?
We at Sigma R&D store methylated DNA frozen.  Cooler temperatures are fine if bacterial contamination does not induce sample degradation.