Non-enzymatic in vitro DNA labeling and label immunoquantification

David Higginson Clarke, Dannelys Pérez-Bello, Ana María Riverón-Rojas, Guillermo Jiménez-Alemán, Mirta Zayas Acosta, Rafaela Pérez Perera, Marquiza Sablón Carrazana, Nelson Díaz Miranda, Chryslaine Rodríguez-Tanty, Esteban Gutiérrez Calzado

Research output: Contribution to journalArticle

1 Citation (Scopus)

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 × 106 bases into thermally denatured DNA were immunoquantified.

Original languageEnglish (US)
Pages (from-to)1-15
Number of pages15
JournalPreparative Biochemistry and Biotechnology
Volume35
Issue number1
DOIs
StatePublished - 2005
Externally publishedYes

Fingerprint

Labeling
Labels
DNA
Nucleotides
Cytidine
Acylation
Antibodies
Chickens
In Vitro Techniques

Keywords

  • Bisulfite-catalyzed transamination reaction
  • Chicken IgY polyclonal antibodies
  • In vitro non-radioactive DNA labeling
  • Label immunoquantification

ASJC Scopus subject areas

  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Biotechnology

Cite this

Clarke, D. H., Pérez-Bello, D., Riverón-Rojas, A. M., Jiménez-Alemán, G., Acosta, M. Z., Perera, R. P., ... Calzado, E. G. (2005). Non-enzymatic in vitro DNA labeling and label immunoquantification. Preparative Biochemistry and Biotechnology, 35(1), 1-15. https://doi.org/10.1081/PB-200041428

Non-enzymatic in vitro DNA labeling and label immunoquantification. / Clarke, David Higginson; Pérez-Bello, Dannelys; Riverón-Rojas, Ana María; Jiménez-Alemán, Guillermo; Acosta, Mirta Zayas; Perera, Rafaela Pérez; Carrazana, Marquiza Sablón; Miranda, Nelson Díaz; Rodríguez-Tanty, Chryslaine; Calzado, Esteban Gutiérrez.

In: Preparative Biochemistry and Biotechnology, Vol. 35, No. 1, 2005, p. 1-15.

Research output: Contribution to journalArticle

Clarke, DH, Pérez-Bello, D, Riverón-Rojas, AM, Jiménez-Alemán, G, Acosta, MZ, Perera, RP, Carrazana, MS, Miranda, ND, Rodríguez-Tanty, C & Calzado, EG 2005, 'Non-enzymatic in vitro DNA labeling and label immunoquantification', Preparative Biochemistry and Biotechnology, vol. 35, no. 1, pp. 1-15. https://doi.org/10.1081/PB-200041428
Clarke DH, Pérez-Bello D, Riverón-Rojas AM, Jiménez-Alemán G, Acosta MZ, Perera RP et al. Non-enzymatic in vitro DNA labeling and label immunoquantification. Preparative Biochemistry and Biotechnology. 2005;35(1):1-15. https://doi.org/10.1081/PB-200041428
Clarke, David Higginson ; Pérez-Bello, Dannelys ; Riverón-Rojas, Ana María ; Jiménez-Alemán, Guillermo ; Acosta, Mirta Zayas ; Perera, Rafaela Pérez ; Carrazana, Marquiza Sablón ; Miranda, Nelson Díaz ; Rodríguez-Tanty, Chryslaine ; Calzado, Esteban Gutiérrez. / Non-enzymatic in vitro DNA labeling and label immunoquantification. In: Preparative Biochemistry and Biotechnology. 2005 ; Vol. 35, No. 1. pp. 1-15.
@article{adc95815b798482694b394de0af720bd,
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 × 106 bases into thermally denatured DNA were immunoquantified.",
keywords = "Bisulfite-catalyzed transamination reaction, Chicken IgY polyclonal antibodies, In vitro non-radioactive DNA labeling, Label immunoquantification",
author = "Clarke, {David Higginson} and Dannelys P{\'e}rez-Bello and River{\'o}n-Rojas, {Ana Mar{\'i}a} and Guillermo Jim{\'e}nez-Alem{\'a}n and Acosta, {Mirta Zayas} and Perera, {Rafaela P{\'e}rez} and Carrazana, {Marquiza Sabl{\'o}n} and Miranda, {Nelson D{\'i}az} and Chryslaine Rodr{\'i}guez-Tanty and Calzado, {Esteban Guti{\'e}rrez}",
year = "2005",
doi = "10.1081/PB-200041428",
language = "English (US)",
volume = "35",
pages = "1--15",
journal = "Preparative Biochemistry and Biotechnology",
issn = "1082-6068",
publisher = "Taylor and Francis Ltd.",
number = "1",

}

TY - JOUR

T1 - Non-enzymatic in vitro DNA labeling and label immunoquantification

AU - Clarke, David Higginson

AU - Pérez-Bello, Dannelys

AU - Riverón-Rojas, Ana María

AU - Jiménez-Alemán, Guillermo

AU - Acosta, Mirta Zayas

AU - Perera, Rafaela Pérez

AU - Carrazana, Marquiza Sablón

AU - Miranda, Nelson Díaz

AU - Rodríguez-Tanty, Chryslaine

AU - Calzado, Esteban Gutiérrez

PY - 2005

Y1 - 2005

N2 - 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 × 106 bases into thermally denatured DNA were immunoquantified.

AB - 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 × 106 bases into thermally denatured DNA were immunoquantified.

KW - Bisulfite-catalyzed transamination reaction

KW - Chicken IgY polyclonal antibodies

KW - In vitro non-radioactive DNA labeling

KW - Label immunoquantification

UR - http://www.scopus.com/inward/record.url?scp=19944430699&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=19944430699&partnerID=8YFLogxK

U2 - 10.1081/PB-200041428

DO - 10.1081/PB-200041428

M3 - Article

C2 - 15704493

AN - SCOPUS:19944430699

VL - 35

SP - 1

EP - 15

JO - Preparative Biochemistry and Biotechnology

JF - Preparative Biochemistry and Biotechnology

SN - 1082-6068

IS - 1

ER -