Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato

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Background: Searching thoroughly for plant cis-elements corresponding to transcription factors is worthwhile to reveal novel gene activation cascades. At the same time, a great deal of research is currently focused on epigenetic events in plants. A widely used method serving both purposes is chromat...

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Autores principales: Ricardi, M.M., González, R.M., Iusem, N.D.
Formato: Artículo publishedVersion
Lenguaje:Inglés
Publicado: 2010
Materias:
Arabidopsis
Lycopersicon esculentum
Solanum
Acceso en línea:http://hdl.handle.net/20.500.12110/paper_17464811_v6_n1_p_Ricardi
Aporte de:
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) de Universidad de Buenos Aires
id paperaa:paper_17464811_v6_n1_p_Ricardi
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spelling paperaa:paper_17464811_v6_n1_p_Ricardi2023-06-12T16:51:18Z Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato Plant Methods 2010;6(1) Ricardi, M.M. González, R.M. Iusem, N.D. Arabidopsis Lycopersicon esculentum Solanum Background: Searching thoroughly for plant cis-elements corresponding to transcription factors is worthwhile to reveal novel gene activation cascades. At the same time, a great deal of research is currently focused on epigenetic events in plants. A widely used method serving both purposes is chromatin immunoprecipitation, which was developed for Arabidopsis and other plants but is not yet operational for tomato (Solanum lycopersicum), a model plant species for a group of economically important crops.Results: We developed a chromatin immunoprecipitation protocol suitable for tomato by adjusting the parameters to optimise in vivo crosslinking, purification of nuclei, chromatin extraction, DNA shearing and precipitate analysis using real-time PCR. Results were obtained with two different antibodies, five control loci and two normalisation criteria.Conclusion: Here we provide a chromatin immunoprecipitation procedure for tomato leaves that could be combined with high-throughput sequencing to generate a detailed map of epigenetic modifications or genome-wide nucleosome positioning data. © 2010 Ricardi et al; licensee BioMed Central Ltd. Fil:Ricardi, M.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:González, R.M. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Iusem, N.D. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2010 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_17464811_v6_n1_p_Ricardi
institution Universidad de Buenos Aires
institution_str I-28
repository_str R-134
collection Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA)
language Inglés
orig_language_str_mv eng
topic Arabidopsis
Lycopersicon esculentum
Solanum
spellingShingle Arabidopsis
Lycopersicon esculentum
Solanum
Ricardi, M.M.
González, R.M.
Iusem, N.D.
Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
topic_facet Arabidopsis
Lycopersicon esculentum
Solanum
description Background: Searching thoroughly for plant cis-elements corresponding to transcription factors is worthwhile to reveal novel gene activation cascades. At the same time, a great deal of research is currently focused on epigenetic events in plants. A widely used method serving both purposes is chromatin immunoprecipitation, which was developed for Arabidopsis and other plants but is not yet operational for tomato (Solanum lycopersicum), a model plant species for a group of economically important crops.Results: We developed a chromatin immunoprecipitation protocol suitable for tomato by adjusting the parameters to optimise in vivo crosslinking, purification of nuclei, chromatin extraction, DNA shearing and precipitate analysis using real-time PCR. Results were obtained with two different antibodies, five control loci and two normalisation criteria.Conclusion: Here we provide a chromatin immunoprecipitation procedure for tomato leaves that could be combined with high-throughput sequencing to generate a detailed map of epigenetic modifications or genome-wide nucleosome positioning data. © 2010 Ricardi et al; licensee BioMed Central Ltd.
format Artículo
Artículo
publishedVersion
author Ricardi, M.M.
González, R.M.
Iusem, N.D.
author_facet Ricardi, M.M.
González, R.M.
Iusem, N.D.
author_sort Ricardi, M.M.
title Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
title_short Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
title_full Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
title_fullStr Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
title_full_unstemmed Protocol: Fine-tuning of a Chromatin Immunoprecipitation (ChIP) protocol in tomato
title_sort protocol: fine-tuning of a chromatin immunoprecipitation (chip) protocol in tomato
publishDate 2010
url http://hdl.handle.net/20.500.12110/paper_17464811_v6_n1_p_Ricardi
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AT gonzalezrm protocolfinetuningofachromatinimmunoprecipitationchipprotocolintomato
AT iusemnd protocolfinetuningofachromatinimmunoprecipitationchipprotocolintomato
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