Radiolabeling Substrates for DNA Repair Assays

Unlabeled nucleotides were purchased from GE Healthcare. [α³²P]-Cordycepin (3′-dATP) and [γ³²P]-ATP were obtained from Perkin Elmer Life Sciences. Substrates were radiolabeled at the 3′ end with [α³²P]-Cordycepin and terminal deoxynucleotidyl transferase (TdT) or at the 5′ end with [γ³²P]-ATP and T4 polynucleotide kinase (T4PNK). TdT, T4PNK, human AP endonuclease I (hAPE1), E. coli Uracil DNA Glycosylase (UDG) and E. coli EndoIII, were from New England Biolabs. Thrombin was obtained from Novagen. BsuLigD was purified as described (19 (link)).

Free full text: Click here

de Ory A., Nagler K., Carrasco B., Raguse M., Zafra O., Moeller R, & de Vega M. (2016). Identification of a conserved 5′-dRP lyase activity in bacterial DNA repair ligase D and its potential role in base excision repair. Nucleic Acids Research, 44(4), 1833-1844.

Publication 2016

Unlabeled nucleotides [γ32P]-ATP E. coli Uracil DNA Glycosylase (UDG)

Cordycepin E coli Endonuclease i Human Nucleotides Polynucleotide kinase Terminal deoxynucleotidyl transferase Thrombin Uracil dna glycosylase

Corresponding Organization : Deutsches Zentrum für Luft- und Raumfahrt e. V. (DLR)

Other organizations : Centro Nacional de Biotecnología

Top 5 similar protocols

Protocol cited in 1 other protocol

Variable analysis

independent variables

Radiolabeling of substrates at the 3' end with [α^32P]-Cordycepin and terminal deoxynucleotidyl transferase (TdT)
Radiolabeling of substrates at the 5' end with [γ^32P]-ATP and T4 polynucleotide kinase (T4PNK)

dependent variables

Not explicitly mentioned

control variables

Unlabeled nucleotides purchased from GE Healthcare
TdT, T4PNK, human AP endonuclease I (hAPE1), E. coli Uracil DNA Glycosylase (UDG) and E. coli EndoIII from New England Biolabs
Thrombin obtained from Novagen
BsuLigD purified as described (19 (link))

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

Based on most similar protocols

Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.

As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!