Alignment Nodes

Alignment nodes map sequencing reads onto a reference. They are typically the most memory- and CPU-intensive step in a pipeline — choose an appropriate resource tier.

BWA-MEM

The standard short-read aligner for DNA (WGS/WES).

Inputs: reads: FASTQ, reference: FASTA Outputs: aligned: BAM

ParameterTypeDefaultDescription
threadsInt4Alignment threads
readGroupString@RG string (required for GATK)
markShortBooltrueMark shorter split hits secondary

Bowtie2

Fast, memory-efficient short-read aligner, popular for ChIP-seq and metagenomics.

Inputs: reads: FASTQ, reference: FASTA Outputs: aligned: BAM

ParameterTypeDefaultDescription
threadsInt4Alignment threads
presetString--sensitiveSensitivity preset

STAR

Splice-aware aligner for RNA-seq. Memory-hungry — building the genome index for a human genome needs ~30 GB+ RAM.

Inputs: reads: FASTQ, reference: FASTA, annotation: GTF Outputs: aligned: BAM, counts: Matrix

ParameterTypeDefaultDescription
threadsInt8Alignment threads
sjdbOverhangInt100readLength - 1 for junction db
quantModeBooltrueEmit gene counts alongside BAM

Post-alignment helpers

NodePurpose
Sort & Index BAMCoordinate-sort and index a BAM (samtools).
Mark DuplicatesFlag PCR/optical duplicates (Picard/GATK).
BAM → CRAMReference-compress alignments to save storage.

Choosing RAM

ReferenceSuggested tier
Bacterial genomeSmall (16 GB)
Human WESMedium (64 GB)
Human WGS / STARLarge (128 GB)+

See also