Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
-The 1.25 M glycine must be at room temperature before use. |
- Use 10,000–300,000 cells or
0.167–5 mg of tissue.
-Whenever possible, use an antibody that is qualified for ChIP.
- Never mix the beads by vortexing; do not allow the beads dry out.
-Keep the formaldehyde incubation time and method consistent between samples.
-Keep the cell lysate cooled on ice during sonication. |
|
Upstream tips |
-The 1.25 M glycine must be at room temperature before use. |
Protocol tips |
- Use 10,000–300,000 cells or
0.167–5 mg of tissue.
-Whenever possible, use an antibody that is qualified for ChIP.
- Never mix the beads by vortexing; do not allow the beads dry out.
-Keep the formaldehyde incubation time and method consistent between samples.
-Keep the cell lysate cooled on ice during sonication. |
Publication protocol
Each ChIP assay used approximately 1.2 × 107 SH-SY5Y cells and was performed with the SOLiD ChIP-Seq Kit (Life Technologies, Foster City, California) according to manufacturer’s specifications, with some adjustments (Supplementary Material, Methods). ChIP-Seq libraries were validated using the BioAnalyzer high-sensitivity chip assay (Agilent, Santa Clara, California) prior to multiplexed high-throughput sequencing on the SOLiD 5500 platform. Fifty-bp single end reads were generated, with a target read number of 25 million tags per sample. In addition to ChIP samples using anti-TCF4 antibodies, their respective IgG controls and input DNA controls were sequenced.
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Manufacturer protocol
Download the product protocol from Thermo Fisher Scientific for SOLiD™ ChIP-Seq Kit, with ChIP magnet below.
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