.. automodule:: oligotag :py:mod:`oligotag` specific options ----------------------------------- .. cmdoption:: -L , --oligo-list= Filename containing a list of oligonucleotides. `oligotag` selects within this list the oligonucleotides that match the specified options. .. CAUTION:: Cannot be used with the ``-s`` option. .. cmdoption:: -s ###, --oligo-size=### Size of oligonucleotides to be generated. .. CAUTION:: Cannot be used with the ``-L`` option. .. WARNING:: A size equal or greater than eight often leads to a very long computing time and requires a large amount of memory. .. cmdoption:: -f ###, --family-size=### Minimal size of the oligonucleotide family to be generated. .. cmdoption:: -d ###, --distance=### Minimal Hamming distance (number of differences) between two oligonucleotides. .. cmdoption:: -g ###, --gc-max=### Maximum number of G or C in the oligonucleotides. .. cmdoption:: -a , --accepted= Selected oligonucleotides are constrained by the given pattern (only :doc:`IUPAC <../iupac>` symbols are allowed). .. CAUTION:: pattern length must have the same length as oligonucleotides. .. cmdoption:: -r , --rejected= Selected oligonucleotides do not match the given pattern (only :doc:`IUPAC <../iupac>` symbols are allowed). .. CAUTION:: pattern length must have the same length as oligonucleotides. .. cmdoption:: -p ###, --homopolymer=### Selected oligonucleotides do not contain any homopolymer longer than the specified length. .. cmdoption:: -P ###, --homopolymer-min=### Selected oligonucleotides contain at least one homopolymer longer or equal to the specified length. .. cmdoption:: -T , --timeout= Timeout to identify a set of oligonucleotides of required size, as defined by the ``-f`` option. .. include:: ../optionsSet/defaultoptions.txt Examples -------- *Example 1:* .. code-block:: bash > oligotag -s 5 -f 24 -d 3 -g 3 -p 2 > mytags.txt Searches for a family of at least 24 oligonucleotides of a length of 5 nucleotides, with at least 3 differences among them, with a maximum of 3 C/G, and without homopolymers longer than 2. The resulting list of oligonucleotides is saved in the ``mytags.txt`` file. *Example 2:* .. code-block:: bash > oligotag -d 5 -L my_oligos.txt -f 10 -p 1 Searches for a subset of at least 10 oligonucleotides listed in the ``my_oligos.txt`` file, with at least 5 differences among them, and without homopolymers. The ``my_oligos.txt`` file must contain a set of oligonucleotides of the same length, with only one oligonucleotide per line. The resulting list of oligonucleotides is printed on the terminal window. *Example 3:* .. code-block:: bash > oligotag -s 7 -f 96 -d 3 -p 1 -r cnnnnnn > mytags.txt Searches for a family of at least 96 oligonucleotides of a length of 7 nucleotides, with at least 3 differences among them, without homopolymers, and without a ``C`` in the first position. The resulting list is saved in the ``mytags.txt`` file. *Example 4:* .. code-block:: bash > oligotag -s 9 -f 24 -d 3 -a yryryryry > mytags.txt Searches for a family of at least 24 oligonucleotides of a length of 9 nucleotides, with at least 3 differences among them, and an alternation of pyrimidines and purines. The resulting list is saved in the ``mytags.txt`` file. Because of the constraints imposed by the ``-a`` option, it is possible to compute longer oligonucleotides in a reasonable time. Reference --------- E. Coissac. Oligotag: a program for designing sets of tags for next-generation sequencing of multiplexed samples. Methods Mol Biol, 888:13-31, 2012.