This tool was developed to convert and compress coverage information from a plain-text tabular pack file (e.g. VG pack). The main goal is to reduce the storage size of the coverage information. It is well integrated into gfa2bin conversion tool for graph-based genome-wide associations studies (GWAS).

Inputs

The general input for the packing is a coverage file in pack format. The format is a tab-separated file with sequence position (seq.pos), node ID (node.id), node offset (node.offset, 0-based) and coverage. An example is shown below.
Sequence-to-graph alignments in GAF and GAM format can be converted to pack format using VG or GAF2PACK. There are several methods to align sequences to graphs, for example VG or GraphAligner. Alternatively you can align to the collection of sequences and inject VG command the linear alignments to the graph.

Example (from data/example_data/9986.1k.txt)

seq.pos	node.id	node.offset	coverage
423	30	61	6
424	30	62	0
425	30	63	2
426	30	64	2
427	31	0	1
428	32	0	1
429	33	0	1
430	33	1	1
431	33	2	0

Output data formats

• pc pack compressed: Compressed representation of a pack file (packing compress).
• pn pack normalized: Compressed representation of pack file after normalization (packing normalize).
• pb pack binary: Represents presence-absence information (from packing bit subcommand).
• pipack index: Index of the graph structure (packing index).

Note

1. The suffix is arbitrary, but helped me to distinguish between the different method outputs.
2. Please consider that the coverage profiles with graphs is very different flat references. Many parts of the graph are not covered at all (see here).

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Install:

git clone https://github.com/MoinSebi/packing
cd packing
cargo build --release

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Usage

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Index

Index a graph or (plain-text) pack file. Index is needed if you want to convert reconvert from pc to pack.

./packing index -g test.gfa -o test.pi 
OR: 
./packing index -p test.pack -o test.pi 

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Compress

Compress a plain-text coverage file to "pack compressed". Mainly used to reduce the storage size of the coverage file. Maximum coverage in these files is 6553. Higher coverages are truncated.

./packing compress -p pack.pack -o pack.pc 

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Conversion methods

General information

Use a threshold to normalize the coverage or create a presence-absence representation.

Thresholds

Absolute threshold:

• -a: A plain number, which will be used as a threshold and is the highest priority.

Dynamic threshold:

• Method -m: Dynamic computation of the threshold based on a method [mean, median, percentile].
• Fraction -f: A relative threshold (fraction) which will be multiplied with the computed value.

Computation
If an absolute threshold is provided, other inputs will be ignored. If a method is provided with -m, we will firstly calculate the specific value (mean or median) which will then scaled by the fraction -f.

Excluding Zeros
Any of these "dynamic" methods can include all entries (default: off, activate with --non-covered) or only the covered entries. The coverage profile on graphs is different compared to flat references, therefore it might be useful to exclude the zeros.

Nodes and sequence If convert your data can either be on sequence and node level, which is also stored in the header of the file. By default we use the sequence based format, but you can change it with the --node flag.

Example computation of threshold
Coverage is: 1, 1, 2, 8, 4, 4
Mean: 4
Fraction: 0.5 Calculated (real) threshold: 2
Normalized coverage (e.g. pc): 0, 0, 1, 4, 2, 2
Binary version (e.g. pt): 0, 0, 1, 1, 1, 1

Default threshold

Without any additional parameters, the default is dynamic threshold with 10% percentile. Values which are equal or above the threshold will be set to 1, all others to 0.

Inputs

• pack Plain-text pack file
• pc Compressed pack
• pnNormalized pack file An index file is needed if you input other than plain-text file for the conversion.

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Bit

Create a presence-absence file (binary, pb) based on a custom threshold.

Example usage

./packing bit -p test.pack -o test.pt -a 5 

On nodes: 
./packing bit -i test.pi -c test.pc --node -o pack.out

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Normalization

Create a normalized coverage file (normalize, pn) based on a custom threshold. Parameters and functionality is similar to the bit subcommand expect that the output is a value-based pack file (normalized).

Example usage

./packing normlaize -p test.pack -o test.pt -a 5  

Include zeros:   
./packing normalize -i test.pi -c test.pc --non-covered -o pack.out 

Additional methods

Info

Information about the index or binary/compressed file. This consists of mostly meta data, e.g. file "type", thresholds and number of entries.

./packing info -i test.pi 
./packing info -c test.pc
./packing info -c test.pt

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View

Show/convert the compressed file in plain text. If the input is a compressed pack (compress output) and an index (see example), you receive a plain-text pack file (comparable with the original pack file). If you don't provide an index, there will be no sequence/node information, just a plain (coverage) vector.

Example usage

./packing view -c test.pc -o test.pc.txt
./packing view -c test.pt -o test.pt.txt
./packing view -c test.pc -i test.pi -o test.pc.full.txt

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Stats

Calculate several stats of pack, pc, pt and pn. Returns information about mean, median, standard deviation and if zeros were removed or not. If the input is sequence level, the output also includes node-level coverage information.

Example usage

./packing stats -p test.pack -o test.packstats
./packing stats -c test.pc -i test.pi -o test.full.stats
./packing stats -c test.pt -o test.pt.stats

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Compare

Compare two pack files. This function is helpful if you want to know if two normalized or presence-absence files have been processed with the same parameter sets.

./packing compare --pack1 test1.pack --pack2 test2.pack

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PC - Pack Compressed - Header explained

Magic bytes explained (in this order):

The header of the file is also compressed (with zstd), therefore you can only read it which packing info or packing view.

Field	Description	Possible values	Bytes
MB	Magic bytes	[35, 38]	2
Sequence	Is sequence	1 (sequence), 0 (node)	1
Keep-zeros	Keep-zeros	1 (yes), 0 (no)	1
PA	DataType	0 = Bit, 1 = Compress, 2 = Normalized	1
Method	Normalization method	0 (Nothing), 1 (Mean), 2(Median), 3(Percentile)	1
fraction	Fraction	Float (f32)	4
Std	Standard deviation multiplier	Float (f32)	4
real_threshold	Real threshold	Float (f32)	4
length	Number of entries	-	4
name	Name of the sample	-	64

In total: 86 bytes

Additional information:

• If method == Nothing but a relative real threshold was set -> Absolute method
• If you are presence/absence, the "real" threshold is enforced: x > threshold
• If the method == Nothing but there is a threshold, it was computed by the "absolute threshold"
• Absolute threshold is always highest priority

TODO

• Node to default in every function