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A simple modification to improve the accuracy of methylation-sensitive restriction enzyme quantitative polymerase chain reaction

Abstrakt

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.

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Kategoria:
Publikacja w czasopiśmie
Typ:
artykuł w czasopiśmie wyróżnionym w JCR
Opublikowano w:
ANALYTICAL BIOCHEMISTRY nr 500, strony 88 - 90,
ISSN: 0003-2697
Język:
angielski
Rok wydania:
2016
Opis bibliograficzny:
Krygier M., Podolak-Popinigis J., Limon J., Sachadyn P., Stanisławska-Sachadyn A.: A simple modification to improve the accuracy of methylation-sensitive restriction enzyme quantitative polymerase chain reaction// ANALYTICAL BIOCHEMISTRY. -Vol. 500, (2016), s.88-90
DOI:
Cyfrowy identyfikator dokumentu elektronicznego (otwiera się w nowej karcie) 10.1016/j.ab.2016.01.020
Weryfikacja:
Politechnika Gdańska

wyświetlono 129 razy

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