Chromatin Immunoprecipitation Assay Protocol

Leaf blades of 60-day-old plants grown in the LD growth chamber were harvested at ZT2 (Two hours after dawn). ChIP assays were performed as described previously using the following antibodies: anti-acetyl-H4K5 (Abcam, Waltham, MA, USA), anti-Flag (Smart-Lifesciences, Changzhou, China), and anti-HA (Smart-Lifesciences, Changzhou, China) antibodies. DNA fragments were recovered by DNA Extraction Reagent (Cat#P1012; shkxbio, Beijing, China) [26 (link)]. Finally, the precipitated DNA was quantified by qPCR with the primers listed in Supplemental Table S1.

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Gong F., Zhang K., Wen J., Yu S., Li W., Du G., Wu C., Zhu K, & Xu Y. (2023). Rice OsMRG702 and Its Partner OsMRGBP Control Flowering Time through H4 Acetylation. International Journal of Molecular Sciences, 24(11), 9219.

Publication 2023

anti-Flag

Antibodies Chip assays Plants leaf Primers

Corresponding Organization : Nanjing Agricultural University

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Variable analysis

independent variables

Growth chamber (LD)
Time of day (ZT2)

dependent variables

DNA fragments precipitated by ChIP assays using anti-acetyl-H4K5, anti-Flag, and anti-HA antibodies

control variables

60-day-old plants
Leaf blades

controls

Positive control: None specified
Negative control: None specified

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