ChIP experiments were performed as previously reported (Zong et al., 2021 (link)). Briefly, 1 g seedlings were used for nuclei isolation. Chromatin was sheared by using a Bioruptor® to an average size of about 500 bp. Sheared chromatin was further diluted in ChIP dilution buffer and incubated with 4 μg (Santa Cruz, c-myc (9E10) X antibody, sc-40X), H3K27me3 (07–449; Millipore Sigma) or GFP (ab290; Abcam) antibodies overnight at 4°C. Antibody was further captured by Dynabeads Protein G (Thermo Fisher Scientific) and then washed by low salt, high salt, LiCl, and TE buffers. The DNA-protein complex was eluted, and reverse cross-linked at 65°C overnight. DNA was purified by using QIAquick® PCR Purification Kit (Qiagen) and was used for qPCR analysis (Primers listed in Table S1 ).
Protocol detail
ChIP-seq protocol for plant chromatin
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Antibodies
Antibody
Buffers
C myc
Chip
Chromatin
Dilution
Isolation
Nuclei
Primers
Protein
Protein g
Salt
Seedlings
Corresponding Organization : The University of Texas at Austin
Top 5 similar protocols
Variable analysis
independent variables
- Chromatin shearing using Bioruptor to an average size of about 500 bp
- DNA-protein complex eluted and reverse cross-linked for qPCR analysis
- 1 g seedlings used for nuclei isolation
- Sheared chromatin further diluted in ChIP dilution buffer
- Incubation with antibodies (c-myc, H3K27me3, GFP) overnight at 4°C
- Antibody captured by Dynabeads Protein G and washed with low salt, high salt, LiCl, and TE buffers
- Positive controls: Not explicitly mentioned
- Negative controls: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!