Copyright (c) 2018,2019 Peter Menzel [email protected]
KIQ is a program for counting the occurrences of a pre-defined static set of k-mers in a large collection of sequencing experiments, for example RNA-Seq.
Occurrence counts for each k-mer per experiment are stored in a database file, which can then be queried for retrieving those experiments that contain a particular k-mer.
KIQ uses 32-mers comprised of the characters A, C, G, and T, which are represented as a 64bit unsigned integer.
git clone https://github.com/pmenzel/kiq
cd kiq/src
make
After compilation, the executable kiq is located in the kiq/bin/ folder.
KIQ can be built with support for directly reading SRA files by using the libraries ncbi-vdb and ngs from NCBI.
To this end, download, compile, and install both of these libraries first:
git clone https://github.com/ncbi/ngs
cd ngs
./configure
make
sudo make install
and
git clone https://github.com/ncbi/ncbi-vdb
cd ncbi-vdb
./configure
make
sudo make install
Afterwards, compile KIQ with
make sra
If the installation of ncbi-vdb and ngs cannot be done with superuser
privileges, then you can specify an existing custom folder as installation
target for ncbi-vdb and ngs when running their ./configure scripts, for
example:
./configure --prefix /home/username/software
In this case, compile KIQ with:
make sra NCBI_DIR=/home/username/software
First, KIQ needs to make an index from a fixed set of k-mers, which are read
line by line from the text file specified with the option -l to the command
kiq index. Additionally, the mandatory options -i and -k are required
for specifying the file names for the k-mer index and the (initially empty)
k-mer count database, which are later used for counting and querying.
kiq index -i kmer_index.bin -k kiq_database.bin -l kmers.txt
Second, the indexed k-mers are counted from sequencing data, either from FASTA/Q or SRA files.
For FASTA/Q files:
kiq db -i kmer_index.bin -k kiq_database.bin -l input.tsv -z 5
The file input.tsv is a tab-separated file, in which the first column denotes
the experiment or sample name/ID and the second column contains the path to a
FASTQ/A file containing the sequence data. Optionally, a third column can
contain a description of the sample, which will also be stored in the database.
For SRA files:
kiq sra -i kmer_index.bin -k kiq_database.bin -l input.tsv -z 5
Here, the second column in the file input.tsv contains the path to a SRA file
associated with the sample. Since KIQ uses the NCBI SRA API, the second column
can also contain just the SRA identifier (starting with SRR). In this case,
the program first checks if the SRA file is already in the local cache and then
reads it from there, or otherwise it will be downloaded on the fly and stored
in the local cache. The cache directory can be set using the
vdb-config tool
from the sra-tools package.
The option -z specifies the number threads that are used for k-mer counting.
Additional datasets can be added to an existing database by using the option -a.
After counting the k-mers, the database can be queried
with kiq query, either by providing a list of query k-mers in a text file
using option -q or by providing query k-mer(s) using option -Q.
For example:
kiq query -i kmer_index.bin -k kiq_database.bin -q query.txt
kiq query -i kmer_index.bin -k kiq_database.bin -Q ACGTACGTACGTACGTACGTACGTACGTACGT
kiq query -i kmer_index.bin -k kiq_database.bin -Q ACGTACGTACGTACGTACGTACGTACGTACGT,GCGTACGTACGTACGTACGTACGTACGTACGG
The k-mer database and metadata can be exported using kiq dump:
kiq dump -i kmer_index.bin -k kiq_database.bin -p stats
kiq dump -i kmer_index.bin -k kiq_database.bin -p metadata
kiq dump -i kmer_index.bin -k kiq_database.bin -p long
kiq dump -i kmer_index.bin -k kiq_database.bin -p stats
KIQ's k-mer database can be modified using kiq modify, which reads a
tab-separated file containing instructions for modifying the database. The
first column contains the experiment ID, the second column contains a command
and the third column contains an optional argument. Two different commands are
available: delete for deleting an experiment (and its associated k-mer
counts) from the database, and update_desc for updating the description for a
given experiment ID.
For example:
SRRZZZ delete
SRRXXX update_desc Total RNA-Seq from Drosophila heads.
Run kiq modify:
kiq modify -i kmer_index.bin -k kiq_database.bin -c modifications.tsv
KIQ uses the BBHash library for indexing of the k-mers and zstr library for reading compressed FASTA/Q files.
See the file LICENSE.