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NAME

       RNAcalibrate    -   calibrate   statistics   of   secondary   structure
       hybridisations of RNAs

SYNOPSIS

       RNAcalibrate [-h] [-d frequency_file] [-f from,to] [-k sample_size] [-l
       mean,std]    [-m    max_target_length]    [-n   max_query_length]   [-u
       iloop_upper_limit] [-v bloop_upper_limit]  [-s]  [-t  target_file]  [-q
       query_file] [target] [query]

DESCRIPTION

       RNAcalibrate  is  a  tool  for  calibrating  minimum  free energy (mfe)
       hybridisations performed with RNAhybrid. It searches a random  database
       that  can  be  given  on the command line or otherwise generates random
       sequences  according  to  given  sample   size,   length   distribution
       parameters  and dinucleotide frequencies. To the empirical distribution
       of length normalised minimum free energies, parameters  of  an  extreme
       value  distribution  (evd)  are fitted. The output gives for each miRNA
       its name (or "command_line" if it was submitted on the  command  line),
       the number of data points the evd fit was done on, the location and the
       scale parameter. The location and scale parameters of the evd can  then
       be given to RNAhybrid for the calculation of mfe p-values.

OPTIONS

       -h     Give a short summary of command line options.

       -d frequency_file
              Generate  random sequences according to dinucleotide frequencies
              given in  frequency_file.  See  example  directory  for  example
              files.

       -f from,to
              Forces  all  structures  to  have  a helix from position from to
              position to with respect  to  the  query.  The  first  base  has
              position 1.

       -k sample_size
              Generate sample_size random sequences. Default value is 5000.

       -l mean,std
              Generate  random  sequences with a normal length distribution of
              mean mean and standard deviation std. Default values are 500 and
              300, respectively.

       -m max_target_length
              The  maximum  allowed  length  of a target sequence. The default
              value is 2000. This option only has an effect if a  target  file
              is given with the -t option (see below).

       -n max_query_length
              The  maximum  allowed  length  of  a query sequence. The default
              value is 30. This option only has an effect if a query  file  is
              given with the -q option (see below).

       -u iloop_upper_limit
              The  maximally  allowed number of unpaired nucleotides in either
              side of an internal loop.

       -v bloop_upper_limit
              The maximally allowed number of unpaired nucleotides in a  bulge
              loop.

       -s     Generate   random   sequences   according  to  the  dinucleotide
              distribution of given targets (either with the -t option  or  on
              command  line. If no -t is given, either the last argument (if a
              -q is given) or the second last argument (if no -q is given)  to
              RNAcalibrate is taken as a target). See -t option.

       -t target_file
              Without   the  -s  option,  each  of  the  target  sequences  in
              target_file is subject to hybridisation with each of the queries
              (which  either are from the query_file or is the one query given
              on command line; see -q below). The sequences in the target_file
              have  to  be in FASTA format, ie. one line starting with a > and
              directly followed by a name, then one or  more  following  lines
              with the sequence itself. Each individual sequence line must not
              have more than 1000 characters.

              With the -s option, the target  (or  target  file)  dinucleotide
              distribution  is  counted,  and  random  sequences are generated
              according to this distribution.

              If no -t is given, random sequences are generated  as  described
              above (see -d option).

       -q query_file
              See  -t  option  above.  If no -q is given, the last argument to
              RNAcalibrate is taken as a query.

REFERENCES

       The energy parameters are taken from:

       Mathews  DH,  Sabina  J,  Zuker  M,  Turner  DH.   "Expanded   sequence
       dependence  of  thermodynamic  parameters  improves  prediction  of RNA
       secondary structure" J Mol Biol., 288 (5), pp 911-940, 1999

VERSION

       This man page documents version 2.0 of RNAcalibrate.

AUTHORS

       Marc Rehmsmeier, Peter Steffen, Matthias Hoechsmann.

LIMITATIONS

       Character dependent energy values are only  defined  for  [acgtuACGTU].
       All other characters lead to values of zero in these cases.

SEE ALSO

       RNAhybrid, RNAeffective