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Enzymatic incorporation and fluorescent labelling of cyclooctyne-modified deoxyuridine triphosphates in DNA
Xiaomei Ren1, Marta Gerowska2, Afaf H El-Sagheer3
1Department of Chemistry, University of Oxford, Chemistry Research Laboratory, Oxford OX1 3TA, UK.
Abstract:
The amino group of 5-aminopropargyl-2'-deoxyuridine-5'-triphosphate was labelled with dibenzocyclooctyne (DIBO) and two derivatives of bicyclo [6.1.0] non-4-yne (BCN) with short and long linkers to produce three different cycloalkyne-modified deoxyuridine triphosphates. BCN was successfully incorporated into DNA at multiple sites by enzyme-mediated primer extension and the polymerase chain reaction (PCR). Efficient fluorescent labelling of the BCN-DNA and DIBO-DNA with Cy3-azide was demonstrated.
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