Home Genetics / Genomics Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells
Genetics / Genomics JoVE (Open Access) Citable · DOI

Automating ChIP-seq Experiments to Generate Epigenetic Profiles on 10,000 HeLa Cells

DOI: 10.3791/52150-v
What you'll learn
  • Automate chromatin immunoprecipitation (ChIP) using robotic liquid handling systems
  • Generate epigenetic profiles from as few as 10,000 cells
  • Reduce labor, cost, and time in ChIP-seq library preparation
  • Map in vivo protein-DNA interactions at scale
Protocol

Methods for mapping in vivo protein-DNA interactions are becoming crucial for every aspect of genomic research but they are laborious, costly, and time consuming. Here a commercially available robotic liquid handling system that automates chromatin immunoprecipitation for mapping in vivo protein-DNA interactions with limited amounts of cells is presented.

Difficulty
advanced
Total time
~2–3 days per batch (automation reduces hands-on time to ~4–6 hours)
Model organism
HeLa cells
Biosafety
BSL-1

Steps

1
Set up robotic chromatin immunoprecipitation workflow

Configure the commercially available liquid handling system for automated ChIP assay. Load samples, reagents, and consumables according to protocol specifications.

▶ 01:40
2
Execute automated library preparation for sequencing

Run the robotic system through chromatin fragmentation, immunoprecipitation, DNA extraction, and library construction steps with minimal manual intervention.

▶ 05:48
3
Analyze generated epigenetic profiles from low cell numbers

Evaluate ChIP-seq results demonstrating successful epigenetic mapping from 10,000 HeLa cells and assess data quality and consistency.

▶ 07:38

🚨 Failure Case Library (3) + Submit your own case

severe
Low Fragmented Chromatin Concentration
DNA concentration of fragmented chromatin preparation is below expected ranges (e.g., <100 µg/ml for HeLa cells, <20 µg/ml for brain tissue). Insufficient material for recommended 5-10 µg chromatin per IP reaction.
💡 4 · ✓ 5
moderate
Low ChIP Signal from Insufficient Starting Material or Antibody
Weak or barely detectable signal across all samples including positive controls. Signal intensity is uniformly low rather than selectively absent at specific regions.
💡 4 · ✓ 4
minor
No Signal at Region of Interest Due to Absent Target
No signal detected at the specific region of interest while ChIP procedure appears technically successful. Other genomic regions or positive controls may show expected signals.
💡 4 · ✓ 4
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