Optimisation of a chromatin immunoprecipitation (ChIP) protocol for histone modification in oil palm


Citation

Norashikin Sarpan, . and Meilina Ong-Abdullah, . and Ooi Siew-Eng, . Optimisation of a chromatin immunoprecipitation (ChIP) protocol for histone modification in oil palm. pp. 242-250. ISSN 1511-2780

Abstract

Chromatin immunoprecipitation (ChIP) is widely used to study protein-DNA interactions such as histone modifications. A ChIP protocol was established for oil palm tissues by adopting ChIP protocols for other plants and optimising several parameters during chromatin extraction DNA shearing and immunoprecipitation. ChIP-DNA was evaluated by end-point polymerase chain reaction (PCR) or quantitative polymerase chain reaction (qPCR). The major modification that improved the yield and quality of DNA obtained at the end of the protocol was that the initial steps were conducted in a cold environment. Frozen tissues were also suitable with this chromatin extraction protocol. Satisfactory results were obtained with the optimised protocol including good quality chromatin and subsequently intact DNA smaller sizes of sheared chromatin and ChIP-enriched DNA suitable for PCR amplification.


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Abstract

Chromatin immunoprecipitation (ChIP) is widely used to study protein-DNA interactions such as histone modifications. A ChIP protocol was established for oil palm tissues by adopting ChIP protocols for other plants and optimising several parameters during chromatin extraction DNA shearing and immunoprecipitation. ChIP-DNA was evaluated by end-point polymerase chain reaction (PCR) or quantitative polymerase chain reaction (qPCR). The major modification that improved the yield and quality of DNA obtained at the end of the protocol was that the initial steps were conducted in a cold environment. Frozen tissues were also suitable with this chromatin extraction protocol. Satisfactory results were obtained with the optimised protocol including good quality chromatin and subsequently intact DNA smaller sizes of sheared chromatin and ChIP-enriched DNA suitable for PCR amplification.

Additional Metadata

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Item Type: Article
AGROVOC Term: Chromatin
AGROVOC Term: Oil palms
AGROVOC Term: Histones
AGROVOC Term: Elaeis guineensis
AGROVOC Term: Gene expression
AGROVOC Term: Isolation
AGROVOC Term: Processing
AGROVOC Term: Cross-linking
AGROVOC Term: Liquid nitrogen
AGROVOC Term: Immunoprecipitation tests
Geographical Term: Malaysia
Depositing User: Ms. Suzila Mohamad Kasim
Last Modified: 28 Apr 2025 06:05
URI: http://webagris.upm.edu.my/id/eprint/24094

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