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PROGRAM:

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.

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