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Sensitive and label-free discrimination of 5-hydroxymethylcytosine and 5-methylcytosine in DNA by ligation-mediated rolling circle amplification

Author:
WangThese authors contributed equally., Zi-yue, Wang, Meng, Zhang, Yan, Zhang, Chun-yang
Source:
Chemical communications 2018 v.54 no.62 pp. 8602-8605
ISSN:
1364-548X
Subject:
DNA, blood serum, fluorescence, oxidation, uracil
Abstract:
We develop a label-free fluorescence method for sensitive discrimination of 5-hydroxymethylcytosine (5hmC) and 5-methylcytosine (5mC) based on the specific oxidation of 5hmC to 5-formylcytosine (5fC) and the subsequent conversion to uracil (U) in combination with ligation-mediated rolling circle amplification (RCA). This method exhibits high sensitivity with a detection limit of as low as 34.8 fM. It can discriminate as low as the 0.01% 5hmC level from the mixture, and exhibits good performance in serum sample analysis as well.
Agid:
6177690