DNA digestion with endonucleases sensitive to CpG methylation such as HpaII followed by polymerase chain reaction (PCR) quantitation is commonly used in molecular studies as a simple and inexpensive solution for assessment of region-specific DNA methylation. We observed that the results of such analyses were highly overestimated if mock-digested samples were applied as the reference.We determined DNA methylation levels in several promoter regions in two setups implementing different references: mock-digested and treated with a restriction enzyme that has no recognition sites within examined amplicons. Fragmentation of reference templates allowed removing the overestimation effect, thereby improving measurement accuracy.
Authors
- Magdalena Krygier,
- dr inż. Justyna Podolak-Popinigis link open in new tab ,
- Prof. dr hab. Janusz Limon,
- prof. dr hab. inż. Paweł Sachadyn link open in new tab ,
- Anna Stanisławska-Sachadyn
Additional information
- DOI
- Digital Object Identifier link open in new tab 10.1016/j.ab.2016.01.020
- Category
- Publikacja w czasopiśmie
- Type
- artykuł w czasopiśmie wyróżnionym w JCR
- Language
- angielski
- Publication year
- 2016
