IFIBYNE   05513
INSTITUTO DE FISIOLOGIA, BIOLOGIA MOLECULAR Y NEUROCIENCIAS
Unidad Ejecutora - UE
artículos
Título:
PROTOCOL: FINE-TUNING OF A CHROMATIN IMMUNOPRECIPITATION (CHIP) PROTOCOL IN TOMATO
Autor/es:
RICARDI MM; GONZÁLEZ RM; IUSEM ND
Revista:
PLANT METHODS
Editorial:
BIOMED CENTRAL LTD
Referencias:
Año: 2010 p. 1 - 8
ISSN:
1746-4811
Resumen:
Abstract 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 parameter 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.