Journal of Oil Palm Research Vol. 30 (2) June 2018, p. 242-250

OPTIMISATION OF A CHROMATIN IMMUNOPRECIPITATION (ChIP) PROTOCOL FOR HISTONE MODIFICATION IN OIL PALM

NORASHIKIN SARPAN*; MEILINA ONG-ABDULLAH* and SIEW-ENG OOI*

DOI: https://doi.org/10.21894/jopr.2018.0021
Published Online: 1 June 2018
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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* Malaysian Palm Oil Board, 6 Persiaran Institusi, Bandar Baru Bangi, 43000 Kajang, Selangor, Malaysia.
E-mail: oseng@mpob.gov.my