title: Non-enzymatic in vitro DNA labeling and label immunoquantification.
abstract: In the present work, a label immunoquantification procedure was developed in order to determine the number of markers introduced into DNA. A non-enzymatic, in vitro labeling method for introducing the p-bromobenzoyl radical (label), through transamination and acylation reactions of the cytidine nucleotides in calf thymus DNA, was carried out. Three spacer arms with different lengths were used for separating the label from the nucleotide and three labeled DNA were obtained. Anti-p-bromobenzoyl chicken IgY polyclonal antibodies were obtained. The antibodies detected the label, into three-labeled DNA, with different sensitivities, in relation to spacer arm length used. About 3-11 labels per 4 x 10(6) bases into thermally denatured DNA were immunoquantified.
authors.affiliation: Molecular Biology Department, Neurosciences Center, Ave. 25, No. 15202, Playa, Havana A.P. 6990, Cuba.
authors.forename: David Higginson
authors.initials: DH
authors.lastname: Clarke
authors.forename: Dannelys
authors.initials: D
authors.lastname: Pérez-Bello
authors.forename: Ana María
authors.initials: AM
authors.lastname: Riverón-Rojas
authors.forename: Guillermo
authors.initials: G
authors.lastname: Jiménez-Alemán
authors.forename: Mirta Zayas
authors.initials: MZ
authors.lastname: Acosta
authors.forename: Rafaela Pérez
authors.initials: RP
authors.lastname: Perera
authors.forename: Marquiza Sablón
authors.initials: MS
authors.lastname: Carrazana
authors.forename: Nelson Díaz
authors.initials: ND
authors.lastname: Miranda
authors.forename: C
authors.initials: C
authors.lastname: Rodríguez-Tanty
authors.forename: Esteban
authors.initials: E
authors.lastname: Gutiérrez Calzado
mesh_terms: D000818:Animals; D002645:Chickens; D004247:DNA / Q000032:analysis* / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D004798:Enzymes / Q000737:chemistry; D015151:Immunoblotting / Q000379:methods*; D007136:Immunoglobulins / Q000032:analysis* / Q000737:chemistry*; D015335:Molecular Probes / Q000032:analysis / Q000737:chemistry; D013194:Staining and Labeling / Q000379:methods*
chemical_list: D004798:Enzymes; C016512:IgY; D007136:Immunoglobulins; D015335:Molecular Probes; D004247:DNA; C427203:calf thymus DNA