Research

Orchestrates the end-to-end germline short-variant pipeline from FASTQ to a filtered, normalized, benchmarked VCF, chaining QC/trim, BWA-MEM2 alignment, duplicate marking, optional BQSR, calling (bcftools/GATK HaplotypeCaller/DeepVariant/DRAGEN), normalization, site+genotype filtering, annotation, and hap.py/vcfeval benchmarking. Use when deciding the pipeline-wide reference-genome commitment (GRCh38 analysis set vs T2T, ALT/decoy handling), sequencing the steps in the defensible order (norma...

Orchestrates the full path from differential expression results to redundancy-collapsed functional enrichment: choose ORA vs GSEA, convert gene IDs per method, run enrichGO/enrichKEGG/enrichPathway/enrichWP or gseGO/gseKEGG (clusterProfiler, ReactomePA, rWikiPathways), and visualize. Use when a DESeq2/edgeR/limma result must become enriched GO terms, KEGG/Reactome/WikiPathways pathways, or a GSEA leading edge; when the input is a full ranking for all genes (GSEA, named decreasing vector) or o...

End-to-end flow, spectral, and mass cytometry (CyTOF) pipeline from raw FCS files to differentially abundant/expressed cell populations. Orchestrates the read -> compensate/unmix -> transform -> QC -> doublet-removal -> cluster-or-gate -> annotate -> diffcyt DA/DS chain with flowCore/CATALYST/diffcyt, branching on instrument type and on clustering-vs-gating. Use when processing a cytometry experiment end-to-end, deciding the pipeline path for an instrument, or wiring the flow-cytometry compon...

Orchestrates an end-to-end CRISPR editing experiment design from target gene to delivery-ready, validatable constructs. Sequences guide design, off-target assessment, edit-modality selection (knockout, base editing, prime editing, HDR knock-in), and template/donor design, with a QC checkpoint at each handoff. Use when designing a complete CRISPR experiment for knockout, point correction, or tagging and the order of operations, the modality decision, and the cross-cutting traps are needed rath...

Classify variant clinical significance with the ACMG/AMP germline framework and its 2018-2025 ClinGen refinements (graded PVS1 decision tree, PM2 downgraded to Supporting, PP5/BP6 retired, calibrated PP3/BP4, Bayesian points), the AMP/ASCO/CAP somatic tiers and ClinGen oncogenicity system, ClinVar star-rating and gnomAD grpmax filtering-AF interpretation. Use when deciding germline-vs-somatic framework, applying current (not flat-2015) ACMG points, checking for a gene-specific VCEP specificat...

Infers directed, time-delayed gene regulatory edges from BULK time-series expression using Granger causality (statsmodels VAR F-test), dynGENIE3 (tree ensembles regressing ODE-derived derivatives; Random Forests by default, Extra-Trees optional), and dynamic Bayesian networks (bnlearn). Use when the output is a RANKED HYPOTHESIS list for perturbation validation, not validated causal edges; deciding Granger vs dynGENIE3 vs DBN by timepoint count and linearity; sizing maxlag against the n>3*max...

Builds draft genome-scale metabolic models from an annotated genome using CarveMe (top-down carving of a BiGG universal model) or gapseq (bottom-up pathway-evidence reconstruction), then loads and sanity-checks the draft in COBRApy. Use when creating a model for an organism without one, choosing between CarveMe and gapseq, gap-filling to a target medium, understanding why a draft that grows is still only a hypothesis, handling BiGG-vs-ModelSEED namespace mismatch, or preparing a draft for cur...

Builds tissue-, cell-type-, and condition-specific metabolic models by integrating transcriptomic or proteomic data into a generic genome-scale model, using extraction algorithms (GIMME, iMAT, INIT/tINIT, MADE, E-Flux, CORDA, FASTCORE) via troppo and corda in Python or the COBRA Toolbox/RAVEN in MATLAB. Use when pruning a generic model to a context, choosing an extraction method and expression threshold, mapping expression through GPR rules to reactions, deciding whether an objective is requi...

Predicts protein and complex structures with deep-learning models (ESMFold, AlphaFold2/ColabFold, AlphaFold3, Chai-1, Boltz-1/2) and reconciles them with confidence metrics. Use when choosing a predictor by input and question rather than novelty (ESMFold single-chain, no-MSA, fast, metagenomic-scale vs AlphaFold3/Chai-1/Boltz for complexes, ligands, nucleic acids, ions, PTMs); recognizing that MSA depth is the dominant accuracy determinant so ESMFold trades accuracy for speed and degrades on ...

Maps protein-protein and protein-ligand interfaces with Bio.PDB, computing contact residues and buried surface area (BSA). Use when choosing a contact cutoff and stating its rationale (heavy-atom 4-5A vs CA-CA 8A vs a SASA-based definition); deciding a contact list is not an interface and computing buried surface area (dSASA/BSA) instead; distinguishing a genuine biological interface from a crystal-packing artifact; identifying ligand-contact or epitope residues; and computing on the biologic...

Detects putative ligand-binding pockets and druggable cavities de novo on an apo protein structure with fpocket, P2Rank, CASTp, and DoGSiteScorer, ranking them by druggability/ligandability score. Use when detecting cavities on an apo structure with no bound ligand; choosing geometric pocket enumeration (fpocket alpha-spheres, CASTp) vs ML ligandability scoring (P2Rank, DoGSiteScorer); recognizing that a geometric cavity is a hypothesis, not automatically a functional or druggable site (may b...

Predicts and prioritizes miRNA target genes with seed-based tools (miRanda, TargetScan, miRDB) and experimentally validated databases (miRTarBase, multiMiR). Use when deciding that a predicted target is a hypothesis not a finding; ranking by the right score (weighted context++, mirSVR, miRDB); raising confidence by intersecting predictions with inversely-correlated mRNA DE; weighing validated (CLIP/reporter) over predicted evidence; or avoiding the circular enrichment of unfiltered target lists.

Trims kit-specific 3' adapters, strips UMIs or 4N degenerate ends, size-selects, and collapses small RNA-seq reads (miRNA, piRNA, tRF) with cutadapt or fastp. Use when choosing the kit's 3' adapter; setting the size window (18-26 nt miRNA vs 24-32 nt piRNA); deciding whether a library carries a true UMI (QIAseq) versus a 4N debiasing spacer (NEXTflex); reading the read-length histogram to judge library quality; or deciding whether to collapse identical reads before mapping.

Quantifies known miRNAs, isomiRs, tRFs, and A-to-I editing fast with miRge3.0 by aligning collapsed reads to curated miRBase or MirGeneDB libraries. Use when choosing miRBase versus MirGeneDB as the reference; deciding whether to collapse isomiRs to the parent miRNA or keep 5'-isomiRs separate (they shift the seed and retarget); confirming the organism is among the six supported species; or remembering that RPM output is for display only and raw counts go to DESeq2/edgeR.

Discovers novel miRNAs and quantifies known miRNAs with miRDeep2 by scoring genome-mapped read stacks against the Dicer/Drosha biogenesis signature. Use when deciding whether a study needs de novo discovery at all versus known-miRNA quantification; choosing the species and related-species miRBase references; reading the miRDeep2 score as a signal-to-noise hypothesis rather than a fixed cutoff; or filtering novel candidates against tRNA/rRNA loci to reject the classic false positives.

Infers developmental trajectories, pseudotime, RNA velocity, and directed fate probabilities from single-cell data using PAGA, Slingshot, Monocle3, DPT, Palantir, scVelo, and CellRank 2. Use when ordering cells along a differentiation continuum, choosing a trajectory method by topology, rooting pseudotime, estimating RNA velocity direction, computing fate probabilities near a bifurcation, or judging whether an inferred trajectory is real.

Detect cluster marker genes and assign manual cell type labels in single-cell RNA-seq using Scanpy (Python) and Seurat (R). Use when finding genes that distinguish clusters, ranking markers for annotation, scoring gene signatures, hand-labeling clusters, or deciding between Wilcoxon marker ranking and pseudobulk condition DE.

Automated reference-based cell type annotation for single-cell RNA-seq using CellTypist, SingleR, Azimuth, scANVI, and scmap to transfer labels from a reference. Use when annotating cell types from a reference atlas or pretrained model, transferring labels onto a query, assessing prediction confidence and rejection, or triaging whether an unexpected cluster is a novel type versus a doublet, low-quality, or batch artifact.

Integrate multiple scRNA-seq samples or batches with Harmony, scVI/scANVI, Seurat (CCA/RPCA), fastMNN, Scanorama, or BBKNN. Resolves which method to use for the dataset size and design, how strongly to correct, when integration is the wrong move (confounded batch/biology), how to score integration with scIB metrics without gaming them, and why corrected expression must not be used for differential expression. Use when integrating batches or datasets, choosing an integration method, diagnosing...

Processes experimental RNA structure probing data (SHAPE-MaP, DMS-MaPseq) into per-nucleotide reactivity profiles with ShapeMapper2, then uses them as soft restraints on thermodynamic folding. Covers reagent and readout choice (SHAPE vs DMS, mutational-profiling vs RT-stop), the three control samples, per-transcript normalization, the Deigan vs Zarringhalam pseudo-energy models, in-cell versus in-vitro interpretation, and multi-conformation deconvolution. Use when converting probing reads to ...

Runs RNA-seq-specific post-alignment QC - strandedness inference, gene-body 5'-3' coverage, read distribution (exonic/intronic/intergenic), rRNA/globin/mitochondrial rate, transcript integrity (TIN), and saturation - with RSeQC, Qualimap, RNA-SeQC, and Picard. Use when validating RNA-seq libraries before quantification or differential expression, diagnosing degradation or gDNA contamination, or determining library strandedness. For raw-FASTQ QC use quality-reports; for UMI dedup use umi-proce...

Aligns RNA-seq reads to a genome with HISAT2, the splice-aware aligner whose hierarchical graph FM-index runs at roughly a quarter of STAR's memory (~7 GB for human), whose SNP/haplotype graph index reduces reference bias in the index itself, and whose MAPQ is GATK-friendly (60 for unique, no 255 problem). Use when RNA alignment must fit a memory-constrained machine, when feeding StringTie/Cufflinks transcript assembly via --dta, or when a SNP-aware graph index is wanted for allele-robust map...

Aligns DNA short reads (paired- or single-end) to a reference genome with bwa-mem2, the maintained successor to BWA-MEM, for WGS/WES and germline/somatic variant-calling pipelines; covers index build, read-group injection, the collate/fixmate/sort/markdup ordering, soft-clipping for SV split reads, ALT/decoy-aware mapping on GRCh38, -K determinism, and streaming straight to a sorted BAM. Use when mapping DNA short reads to a reference for variant calling, coverage, ChIP/ATAC (alongside bowtie...

Aligns DNA short reads to a reference with Bowtie2, choosing end-to-end (whole read must align) vs local (soft-clip read ends) mode and a sensitivity preset; the de-facto aligner for ChIP-seq, ATAC-seq, and CUT&RUN, where fragment-geometry flags (--no-mixed, --no-discordant, --dovetail, -X) and a tool-appropriate MAPQ filter feed the peak caller. Use when aligning ChIP/ATAC/CUT&RUN reads, when read ends are adapter-contaminated and need soft-clipping, or when a tunable sensitivity/speed prese...