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

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

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 amp...

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Veröffentlicht in:Chemical communications (Cambridge, England) England), 2018, Vol.54 (62), p.8602-8605
Hauptverfasser: Wang, Zi-Yue, Wang, Meng, Zhang, Yan, Zhang, Chun-Yang
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Sprache:eng
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Amplification
Deoxyribonucleic acid
Discrimination
DNA
Fluorescence
Oxidation
Uracil
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Wang, Meng
Zhang, Yan
Zhang, Chun-Yang
description 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.
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source Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection
subjects Amplification
Deoxyribonucleic acid
Discrimination
DNA
Fluorescence
Oxidation
Uracil
title Sensitive and label-free discrimination of 5-hydroxymethylcytosine and 5-methylcytosine in DNA by ligation-mediated rolling circle amplification
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