ZNA (Compressed Z-Nucleic N-Acid A) is a high-performance binary format for storing DNA/RNA sequences with exceptional compression and I/O speed.
- 135 MB/s roundtrip throughput (9.5x faster than Python baseline)
- 2.8+ GB/s encoding/decoding for long reads
- 3.7-4.0x compression ratio with Zstd
- C++ acceleration with pure Python fallback
- High Compression: 2-bit encoding (4 bases per byte) + optional Zstd compression
- Ultra-Fast I/O: C++ accelerated encode/decode with block-based architecture
- Minimal Dependencies:
zstandardonly (C++ extension auto-compiled) - Flexible: Single-end, paired-end, and interleaved reads
- Strand-Specific Support: dUTP, TruSeq, and custom strand protocols
- Built-in Shuffle: Memory-bounded random shuffling for training data preparation
- Metadata Rich: Read groups, descriptions, and custom flags
- Unix-Friendly: Pipe-compatible CLI for seamless workflow integration
- Streaming: Memory-efficient block-based processing
# From source (recommended - includes C++ acceleration)
git clone https://github.com/mkiyer/zna.git
cd zna
pip install -e .
# Check if C++ acceleration is available
python -c "from zna.core import is_accelerated; print(f'Accelerated: {is_accelerated()}')"Requirements:
- Python ≥3.10
- C++ compiler (for optimal performance)
- CMake ≥3.15 (auto-installed via pip)
# Encode FASTQ to compressed ZNA (default: Zstd level 3)
zna encode sample.fastq.gz -o sample.zna
# Encode with shuffle (for ML training data)
zna encode sample.fastq.gz --shuffle -o shuffled.zna
# Encode with shuffle and explicit memory cap per bucket
zna encode sample.fastq.gz --shuffle --shuffle-buffer-size 512M -o shuffled.zna
# Shuffle an existing ZNA file
zna shuffle input.zna -o shuffled.zna
# Decode back to FASTA
zna decode sample.zna -o sample.fasta
# Inspect file statistics
zna inspect sample.zna
# Pipe-friendly workflows
cat reads.fastq | zna encode -o reads.zna
zna decode reads.zna | head -n 1000| Read Type | Encode (MB/s) | Decode (MB/s) | Compression |
|---|---|---|---|
| Short (Illumina, 100-150bp) | 189.5 | 668.8 | 3.68x |
| Medium (300-500bp) | 540.5 | 1,280.9 | 3.87x |
| Long (PacBio, 1-5kb) | 1,921.5 | 2,864.6 | 3.98x |
| Very Long (Nanopore, 5-15kb) | 2,824.7 | 3,392.7 | 3.99x |
Key Insights:
- Performance scales dramatically with read length
- Compression ratio remains consistent across workloads
- C++ acceleration provides 9.5x speedup over pure Python
See docs/PERFORMANCE.md for detailed benchmarking.
- docs/RELEASING.md - Publishing to PyPI and Bioconda
- docs/PERFORMANCE.md - Benchmarks and tuning
ZNA files use a binary format optimized for nucleic acid sequences:
- File Extension:
.zna(for both compressed and uncompressed files) - Default Compression: Zstd level 3 (use
--uncompressedflag to disable) - Magic Number:
ZNA\x1A(4 bytes) - Version: 1 (1 byte)
- 2-bit Encoding: A=00, C=01, G=10, T=11
- Block Structure: Data organized in compressed/uncompressed blocks
- Metadata: Read groups, descriptions, and custom information
┌─────────────────────────────────────┐
│ File Header │
│ - Magic (4 bytes) │
│ - Version (1 byte) │
│ - Sequence length encoding (1 byte)│
│ - Flags (1 byte) │
│ - Compression method (1 byte) │
│ - Compression level (1 byte) │
│ - Metadata lengths (6 bytes) │
│ - Variable metadata strings │
├─────────────────────────────────────┤
│ Block 1 │
│ - Block Header (12 bytes) │
│ * Compressed size (4 bytes) │
│ * Uncompressed size (4 bytes) │
│ * Record count (4 bytes) │
│ - Compressed/Raw Payload │
│ * Record 1: flags, length, seq │
│ * Record 2: flags, length, seq │
│ * ... │
├─────────────────────────────────────┤
│ Block 2 │
│ ... │
└─────────────────────────────────────┘
Each record in a block contains:
- Flags (1 byte): IS_READ1, IS_READ2, IS_PAIRED
- Length (1-4 bytes): Sequence length (configurable)
- Sequence (variable): 2-bit encoded bases
- Method 0: Uncompressed (
.zna) - Method 1: Zstd compression (
.zzna, levels 1-22) - Block Size: Default 128KB (configurable)
# From FASTQ file
zna encode sample.fastq -o sample.zna
# From FASTA file
zna encode sample.fasta -o sample.zna
# From gzipped input
zna encode sample.fastq.gz -o sample.zzna
# With high compression (default is level 3)
zna encode sample.fastq --level 5 -o sample.zna
# Uncompressed (rarely needed)
zna encode sample.fastq --uncompressed -o sample.zna
# From stdin
cat sample.fastq | zna encode -o sample.zna
# Force format (when extension detection fails)
cat data.txt | zna encode --fastq -o sample.zna# Separate R1/R2 files
zna encode R1.fastq.gz R2.fastq.gz -o paired.zna
# Interleaved file (strict alternating R1/R2 pairs)
zna encode interleaved.fastq --interleaved -o paired.zna
# Interleaved from stdin
cat interleaved.fastq | zna encode --interleaved -o paired.zznaThe --interleaved mode intelligently detects both paired-end and single-end reads in the same file by analyzing read names. This is useful for output from tools like fastp that produce mixed merged (single) and unmerged (paired) reads.
How it works:
- Reads with matching base names (e.g.,
read1/1andread1/2) are paired - Reads without matching pairs are treated as single-end
- Read names are used to determine pairing (not just alternating order)
# Mixed interleaved input (fastp output with merged + unmerged reads)
zna encode fastp_output.fastq --interleaved -o mixed.zna
# Example input structure:
# @read1/1 → paired with next read
# @read1/2
# @merged1 → single-end (no pair)
# @read2/1 → paired with next read
# @read2/2
# @merged2 → single-end (no pair)Read name formats supported:
/1and/2suffixes:read1/1,read1/2- No suffix: treated as single-end unless next read has matching base name
- Comments ignored:
read1/1 merged_length:150extractsread1/1
Strand normalization of merged/single reads: single-end reads (including merged
reads with no mate) are treated as read1 for strand normalization. Under
--strand-specific, a single read is reverse-complemented exactly when read1 is
antisense, so merged reads end up on the same strand as normalized paired R1 reads.
# Custom metadata
zna encode sample.fastq \
--read-group "Sample_01" \
--description "Experiment XYZ" \
-o sample.zna
# Strand-specific library (default: R1 antisense, R2 sense)
zna encode R1.fastq.gz R2.fastq.gz \
--strand-specific \
-o stranded.zna
# Custom strand orientation (e.g., fr-secondstrand protocol)
zna encode R1.fastq.gz R2.fastq.gz \
--strand-specific --read1-sense --read2-antisense \
-o stranded.zna
# Handle sequences with N nucleotides
zna encode sample.fastq --npolicy drop -o clean.zna # Skip sequences with N
zna encode sample.fastq --npolicy random -o clean.zna # Replace N with random base
zna encode sample.fastq --npolicy A -o clean.zna # Replace N with A
# Shuffle during encoding (for ML training data preparation)
zna encode sample.fastq --shuffle -o shuffled.zna
zna encode R1.fastq.gz R2.fastq.gz --shuffle --seed 12345 -o shuffled.zna
# Control compression
zna encode sample.fastq \
--level 9 \
--block-size 262144 \
-o sample.zna
# Uncompressed (rarely needed, for maximum I/O speed)
zna encode sample.fastq --uncompressed -o sample.zna
# Sequence length encoding (max sequence length)
zna encode sample.fastq \
--seq-len-bytes 1 \ # Max 255 bp
-o short_reads.zna
zna encode sample.fastq \
--seq-len-bytes 2 \ # Max 65,535 bp (default)
-o sample.zna
zna encode sample.fastq \
--seq-len-bytes 4 \ # Max 4.2 billion bp
-o long_reads.zna# To FASTA file
zna decode sample.zna -o output.fasta
# To gzipped FASTA
zna decode sample.zna -o output.fasta.gz
# To stdout (pipe-friendly)
zna decode sample.zna | head -n 1000
# From stdin
cat sample.zna | zna decode -o output.fasta# Interleaved output (default)
zna decode paired.zna -o interleaved.fasta
# Split to R1/R2 files (use # placeholder)
zna decode paired.zna -o reads#.fasta
# Creates: reads_1.fasta and reads_2.fasta
# Split with gzip
zna decode paired.zna -o reads#.fasta.gz
# Creates: reads_1.fasta.gz and reads_2.fasta.gz
# Restore original strand for strand-specific libraries
zna decode stranded.zna --restore-strand -o reads.fasta# Extract first 1M reads
zna decode large.zna | head -n 2000000 > subset.fasta
# Count sequences
zna decode sample.zna | grep -c "^>"
# Convert to gzipped output via pipe
zna decode sample.zna --gzip > output.fasta.gz
# Chain operations
zna decode sample.zna | seqtk seq -r - | gzip > reversed.fasta.gz# Show file statistics
zna inspect sample.znaExample Output:
File: sample.zna
Total Size: 45.32 MB
--- Header Metadata ---
Read Group: Sample_01
Description: Experiment XYZ
Seq Length: 2 bytes (Max: 65535 bp)
Strand Specific: True
R1 Antisense: True
R2 Antisense: False
Compression: ZSTD (Level 3)
--- Content Statistics ---
Total Blocks: 356
Total Records: 1000000
Compressed Payload: 42.15 MB
Uncompressed Data: 125.50 MB
Compression Ratio: 2.98x
Convert FASTQ/FASTA to ZNA format.
Usage:
zna encode [FILE1] [FILE2] [OPTIONS]
Positional Arguments:
FILE1 [FILE2] Input files (0=stdin, 1=single/interleaved, 2=paired R1 R2)
Options:
--interleaved Treat input as interleaved (auto-detects mixed paired/single reads)
--shuffle Shuffle records after encoding (for ML training data)
--seed N Random seed for --shuffle (default: 42)
--shuffle-buffer-size N
Max memory per bucket for encode --shuffle (default: 1G).
Accepts K/M/G suffixes.
--fasta Force FASTA format (overrides extension detection)
--fastq Force FASTQ format (overrides extension detection)
Metadata:
--read-group TEXT Read group ID (default: "Unknown")
--description TEXT Description string
--strand-specific Flag library as strand-specific (default: R1 antisense, R2 sense)
--strand-normalize Enable strand normalization (RC reads to consistent strand).
With --strand-specific: deterministic (antisense reads RC'd).
Without: random RC (for unstranded data).
--read1-sense Read 1 represents sense strand
--read1-antisense Read 1 represents antisense strand (default when --strand-specific)
--read2-sense Read 2 represents sense strand (default when --strand-specific)
--read2-antisense Read 2 represents antisense strand
--npolicy {drop,random,A,C,G,T}
Policy for handling 'N' nucleotides:
- drop: skip sequences containing N
- random: replace N with random base (A/C/G/T)
- A/C/G/T: replace N with specific base
Format Options:
-o, --output FILE Output file (default: stdout)
--seq-len-bytes N Bytes for sequence length: 1, 2, or 4 (default: 2)
--block-size N Block size in bytes (default: 131072)
--zstd Force Zstd compression
--uncompressed Force uncompressed
--level N Zstd compression level 1-22 (default: 3)
Convert ZNA to FASTA format.
Usage:
zna decode [FILE] [OPTIONS]
Positional Arguments:
FILE Input ZNA file (default: stdin)
Options:
-o, --output FILE Output FASTA file. Use '#' for split R1/R2
-q, --quiet Suppress progress messages
--gzip Force gzip compression for stdout
--restore-strand Restore original strand orientation for antisense reads
Display ZNA file statistics.
Usage:
zna inspect FILE [--counts]
input FILE Input ZNA file to inspect
--counts Also report per-flag record counts (paired R1, paired R2,
single/merged, reverse-complemented). Reads block payloads,
so slower than the default header-only scan.
Randomly shuffle records in a ZNA file with bounded memory usage. Preserves paired-end read associations.
Usage:
zna shuffle INPUT -o OUTPUT [OPTIONS]
Positional Arguments:
INPUT Input ZNA file to shuffle
Options:
-o, --output FILE Output ZNA file (required)
-s, --seed N Random seed for reproducibility (default: 42)
-b, --buffer-size SIZE Maximum memory per bucket (default: 1G)
Accepts K/M/G suffixes (e.g., 512M, 2G)
--block-size SIZE Block size for output ZNA (default: 4M)
--tmp-dir DIR Directory for temporary files (default: system temp)
-q, --quiet Suppress progress messages
Algorithm: Uses bucket shuffle with bounded memory:
- Randomly distributes records into K temporary bucket files on disk
- Shuffles each bucket in memory using Fisher-Yates algorithm
- Concatenates shuffled buckets to produce uniform random permutation
Examples:
# Shuffle with default settings (1GB memory, seed 42)
zna shuffle input.zna -o shuffled.zna
# Shuffle with custom seed for reproducibility
zna shuffle input.zna -o shuffled.zna --seed 12345
# Shuffle with limited memory (512MB buffer)
zna shuffle input.zna -o shuffled.zna --buffer-size 512M
# Shuffle paired-end data (pairs stay together)
zna shuffle paired.zna -o shuffled_paired.znaNote: Paired-end reads (R1+R2) are kept together as a single shuffle unit.
Typical compression ratios compared to raw FASTQ:
| Format | Size | Ratio | Notes |
|---|---|---|---|
| FASTQ (uncompressed) | 100% | 1.0x | Baseline |
| FASTQ.gz (gzip -6) | 25-30% | 3-4x | Standard |
| ZNA (uncompressed) | 12-15% | 6-8x | 2-bit encoding only |
| ZNA (Zstd L3) | 8-10% | 10-12x | Fast compression (default) |
| ZNA (Zstd L9) | 6-8% | 12-16x | High compression |
Results vary based on sequence complexity and redundancy
- Encoding: ~5-10M reads/second (single thread)
- Decoding: ~8-15M reads/second (single thread)
- Block-based: Enables parallel processing (future)
- Streaming I/O: Constant memory usage
- Default block size: 128KB buffer
- No index required: Sequential scan
DNA bases are encoded in 2 bits:
A = 00 = 0
C = 01 = 1
G = 10 = 2
T = 11 = 3
Four bases pack into one byte:
Byte: [B1][B2][B3][B4]
76 54 32 10 (bit positions)
Pre-computed lookup tables provide O(1) encoding/decoding:
- Encoding: 256-element array mapping ASCII → 2-bit
- Decoding: 256-element tuple mapping byte → 4-character string
Data is organized in independently compressed blocks:
- Advantages: Random access, parallel processing potential
- Overhead: ~12 bytes per block
- Optimal size: 128KB balances compression ratio and I/O
- Zstd: Modern compression algorithm (Facebook)
- Reusable compressor: Amortizes initialization cost
- Memoryview parsing: Zero-copy decompression
- Pre-sized buffers: Eliminates reallocations
ZNA supports strand-specific RNA-seq libraries by normalizing all reads to sense strand orientation during encoding. This enables consistent downstream analysis while preserving the ability to restore original strand information.
- Encoding: Antisense reads are reverse-complemented to sense strand
- Storage: All reads stored in sense orientation
- Decoding: Use
--restore-strandto recover original orientation
The --strand-normalize flag controls whether reads are reverse-complemented
to a consistent strand during encoding:
- With
--strand-specific: Deterministic normalization — antisense reads are reverse-complemented to sense orientation based on the library protocol. Each read's IS_RC flag records whether it was flipped. - Without
--strand-specific: Random reverse-complementing for unstranded data. Useful for data augmentation in ML training. - Without
--strand-normalize: Reads are stored in their original orientation (no reverse-complementing).
# Strand-normalized encoding (most common for stranded RNA-seq)
zna encode R1.fq.gz R2.fq.gz --strand-specific --strand-normalize -o lib.zna
# Decode with original strand orientation restored
zna decode lib.zna --restore-strand -o original.fasta
# Decode with sense-normalized sequences (for alignment)
zna decode lib.zna -o normalized.fasta| Flag | Description |
|---|---|
--strand-specific |
Enable strand-specific mode (default: R1 antisense, R2 sense) |
--read1-sense |
Read 1 represents sense strand |
--read1-antisense |
Read 1 represents antisense strand |
--read2-sense |
Read 2 represents sense strand |
--read2-antisense |
Read 2 represents antisense strand |
| Protocol | R1 | R2 | ZNA Flags |
|---|---|---|---|
| dUTP / TruSeq Stranded | antisense | sense | --strand-specific (default) |
| Illumina Stranded mRNA | antisense | sense | --strand-specific |
| fr-firststrand | antisense | sense | --strand-specific |
| fr-secondstrand | sense | antisense | --strand-specific --read1-sense --read2-antisense |
| Ligation (ScriptSeq) | sense | antisense | --strand-specific --read1-sense --read2-antisense |
# dUTP/TruSeq protocol (most common - this is the default)
zna encode R1.fastq.gz R2.fastq.gz --strand-specific -o library.zzna
# fr-secondstrand protocol
zna encode R1.fastq.gz R2.fastq.gz \
--strand-specific --read1-sense --read2-antisense \
-o library.zzna
# Decode with sense-normalized sequences (for alignment)
zna decode library.zzna -o normalized.fasta
# Decode with original strand orientation restored
zna decode library.zzna --restore-strand -o original.fastaZNA can store numeric metadata as compact columnar label columns alongside
each sequence. Labels are parsed from key-value tags in FASTQ headers
(e.g. output from samtools fastq -T), but the tag format is not limited
to SAM — any KEY:TYPE:VALUE field in the header will work, and keys
can be any length.
# Two labels with descriptions
zna encode reads.fq.gz -o reads.zna \
--label NH:C --label AS:i \
--label-desc NH:"Number of hits" --label-desc AS:"Alignment score"The --label format is NAME:TYPE where TYPE is one of:
A, c, C, s, S, i, I, f, d, q, Q.
Use the smallest type that fits your data to minimize file size.
By default, the label name (stored in the ZNA header) is also used as
the tag to parse from input. You can decouple these with the
3-part format NAME:TYPE:TAG:
# Store as "edit_dist" in ZNA, but parse "NM" tag from input headers
zna encode reads.fq.gz -o reads.zna \
--label edit_dist:C:NM --label aln_score:i:AS
# Custom long-form tags (not SAM) work too
zna encode reads.fq.gz -o reads.zna \
--label score:i:alignment_score --label edits:C:edit_distanceThe tag is only used at encode time and is not stored in the ZNA file.
When decoding, the label name is used in the output.
For many labels, define them in a YAML file instead of many CLI flags:
zna encode reads.fq.gz -o reads.zna --label-defs labels.yaml# labels.yaml
labels:
- name: NM
type: C
description: Edit distance
missing: 255
- name: aln_score
type: i
tag: AS # parse "AS" from input, store as "aln_score"
description: Alignment score
missing: -1CLI flags --label and --label-desc override values from the YAML
file, so you can keep a YAML base and tweak individual labels per run.
See examples/labels.yaml for a fully-commented
template.
# Include labels as SAM-style tags in the output
zna decode reads.zna --labels > output.fq
# Inspect to see label definitions
zna inspect reads.znafrom zna.core import ZnaHeader, ZnaWriter, ZnaReader
from zna.dtypes import LabelDef, parse_dtype
defs = (
LabelDef(0, "NM", "Edit distance", parse_dtype("C"), missing=255),
LabelDef(1, "AS", "Alignment score", parse_dtype("i"), missing=-1),
)
header = ZnaHeader(read_group="sample", labels=defs)
with open("out.zna", "wb") as f:
with ZnaWriter(f, header) as w:
w.write_record("ACGT", is_paired=False,
is_read1=False, is_read2=False,
label_values=(3, 280))
with open("out.zna", "rb") as f:
reader = ZnaReader(f)
for seq, is_paired, is_r1, is_r2, labels in reader.records_with_labels():
print(seq, labels) # ACGT (3, 280)- ✅ Long-term archival: High compression with fast retrieval
- ✅ Data transfer: Reduced bandwidth requirements
- ✅ Cloud storage: Lower storage costs
- ✅ Pipeline integration: Unix-friendly streaming
- ✅ Reference storage: Efficient genome/transcriptome storage
- ❌ Random access: Sequential format (no index)
- ❌ Quality scores: Sequences only (use CRAM/BAM for qualities)
- ❌ Small files: Overhead outweighs benefits (<10K reads)
- ❌ Real-time streaming: Use case requires quality scores
| Feature | ZNA | FASTA | FASTQ | CRAM | FASTA.gz |
|---|---|---|---|---|---|
| Compression | Excellent | None | None | Excellent | Good |
| Speed | Fast | Fastest | Fast | Slow | Medium |
| Quality Scores | ❌ | ❌ | ✅ | ✅ | ❌ |
| Paired-End | ✅ | ❌ | ❌ | ✅ | ❌ |
| Random Access | ❌ | ✅ | ✅ | ✅ | ❌ |
| Streaming | ✅ | ✅ | ✅ | Limited | ✅ |
| Dependencies | 1 | 0 | 0 | Many | 0 |
In addition to the CLI, ZNA provides a Python API:
from zna import ZnaHeader, ZnaWriter, ZnaReader, COMPRESSION_ZSTD
# Writing
header = ZnaHeader(
read_group="Sample_01",
compression_method=COMPRESSION_ZSTD,
compression_level=5
)
with open("output.zzna", "wb") as f:
with ZnaWriter(f, header) as writer:
writer.write_record("ACGTACGT", is_paired=False,
is_read1=False, is_read2=False)
writer.write_record("TGCATGCA", is_paired=False,
is_read1=False, is_read2=False)
# Reading
with open("output.zzna", "rb") as f:
reader = ZnaReader(f)
print(f"Read Group: {reader.header.read_group}")
for seq, is_paired, is_read1, is_read2 in reader.records():
print(seq)# All tests
PYTHONPATH=src pytest -v
# Specific test suite
PYTHONPATH=src pytest tests/test_cli.py -v
PYTHONPATH=src pytest tests/test_core.py -v
# With coverage
PYTHONPATH=src pytest --cov=zna tests/# Format code
black src/ tests/
# Type checking
mypy src/zna/- Sequences only: No quality scores, headers, or annotations
- Sequential access: No random access without full scan
- DNA/RNA only: A, C, G, T bases (N or IUPAC codes not supported)
- Case insensitive: Lowercase converted to uppercase
- No index: Full file scan required for record counting
- Parallel compression/decompression
- Optional index for random access
- Support for IUPAC ambiguity codes
- Memory-mapped I/O for large files
- Streaming statistics (GC content, length distribution)
GNU General Public License v3.0 (GPLv3)
If you use ZNA in your research, please cite:
Iyer, M. (2026). ZNA: A compressed binary format for nucleic acid sequences.
GitHub: https://github.com/mkiyer/zna
Contributions are welcome! Please:
- Fork the repository
- Create a feature branch
- Add tests for new functionality
- Ensure all tests pass
- Submit a pull request
- Author: Matthew Iyer
- Email: mkiyer@umich.edu
- Issues: https://github.com/mkiyer/zna/issues