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Example Workflow Prompts

This guide provides example prompts for different workflow types in FlowAgent. These prompts demonstrate how to effectively communicate your analysis requirements to the system.

RNA-seq Analysis

Basic Differential Expression

Analyze differential expression in RNA-seq data from breast cancer samples vs controls. I have:
- Paired-end FASTQ files in data/raw/
- Sample metadata in metadata.csv with condition labels
- Human genome reference (hg38)
Please include:
- Quality control with FastQC
- DESeq2 normalization
- Pathway analysis
- Volcano plot visualization

Single-cell Analysis

Process single-cell RNA-seq data from 10x Genomics:
- Cell Ranger output in scRNA/counts/
- 8 samples (4 treated, 4 control)
- Human reference (GRCh38)
Analysis needed:
- Seurat preprocessing and QC
- Integration of all samples
- Differential expression by cluster
- Trajectory analysis with Monocle3

ChIP-seq Analysis

Histone Modification

Process ChIP-seq data for H3K27ac marks in neural progenitor cells. I have:
- ChIP FASTQ files in chip/
- Input control in input/
- mm10 genome
Requirements:
- Remove duplicates and low MAPQ reads
- Call peaks with MACS2
- Find enriched motifs
- Generate coverage plots around TSS regions

Transcription Factor Binding

Analyze ChIP-seq for CTCF binding:
- Raw reads in tf_chip/fastq/
- Matched input controls
- hg38 genome
Include:
- IDR analysis between replicates
- Motif discovery with MEME
- Conservation analysis
- Integration with Hi-C data

Hi-C Analysis

Basic Interaction Analysis

Analyze chromosome interactions in Hi-C data from fibroblasts. Files:
- Raw reads in hic/reads/
- hg38 reference genome
- Restriction enzyme: MboI
Analysis needs:
- Generate contact matrices at multiple resolutions
- Call TADs and loop domains
- Find significant interactions
- Compare with published compartment annotations

Multi-condition Comparison

Compare Hi-C data between wild-type and knockout:
- WT data in hic/wt/
- KO data in hic/ko/
- mm10 genome, DpnII digestion
Required:
- Matrix normalization
- Differential interaction analysis
- TAD boundary comparison
- Integration with RNA-seq changes

ATAC-seq Analysis

Basic Accessibility

Run ATAC-seq analysis on T cell activation data. Input:
- FASTQ files in atac/fastq/
- Sample groups: resting vs activated
- Human genome GRCh38
Required analysis:
- Fragment size distribution QC
- Call accessible regions
- Find differential accessibility
- Identify TF footprints
- Integrate with RNA-seq from matching samples

Time Series Analysis

Process ATAC-seq time series during differentiation:
- Samples from 0h, 12h, 24h, 48h
- Technical replicates in atac/timeseries/
- Mouse genome mm10
Analysis:
- Quality metrics across time points
- Accessibility dynamics
- TF motif enrichment changes
- Pseudotime ordering of regions

Best Practices for Prompts

When creating prompts for FlowAgent, consider including:

  1. Data Description
  2. File locations and formats
  3. Sample organization
  4. Reference genome
  5. Experimental design

  6. Analysis Requirements

  7. Quality control steps
  8. Core analysis methods
  9. Statistical approaches
  10. Integration needs

  11. Output Expectations

  12. Required visualizations
  13. File formats
  14. Statistical thresholds
  15. Validation metrics

  16. Additional Context

  17. Biological background
  18. Previous findings
  19. Related datasets
  20. Publication requirements

Using Custom Scripts

You can request specific custom scripts in your prompts:

Please run RNA-seq analysis with:
- Custom normalization script (deseq2_normalize)
- Custom QC metrics (rna_qc_extended)
- Standard alignment and quantification

Combining Workflows

For multi-omic analysis, you can combine workflows:

Integrate ATAC-seq and RNA-seq data:
- ATAC data in atac/fastq/
- RNA data in rna/fastq/
- Matching time points and conditions
Analysis:
- Process each assay independently
- Correlate accessibility with expression
- Find coordinated changes
- Generate integrated regulatory networks