| dan

## Function

Calculates DNA RNA/DNA melting temperature

## Description

Dan calculates the melting temperature (Tm) and the percent G+C of a nucleic acid sequence (optionally plotting them). For the Melting temperature profile, free energy values calculated from nearest neighbor thermodynamics are used (Breslauer et al. Proc. Natl. Acad. Sci. USA 83, 3746-3750 and Baldino et al. Methods in Enzymol. 168, 761-777).

## Usage

Here is a sample session with dan

    % dan
    Calculates DNA RNA/DNA melting temperature
    Input sequence(s): tembl:paamir
    Enter window size [20]:
    Enter Shift Increment [1]:
    Enter DNA concentration (nM) [50.]:
    Enter salt concentration (mM) [50.]:
    Output report [paamir.dan]:

Go to the input files for this example
Go to the output files for this example

Example 2

An example of producing a plot of Tm:

    % dan -plot -graph cps
    Calculates DNA RNA/DNA melting temperature
    Input sequence(s): tembl:paamir
    Enter window size [20]:
    Enter Shift Increment [1]:
    Enter DNA concentration (nM) [50.]:
    Enter salt concentration (mM) [50.]:
    Enter minimum temperature [55.]:

    Created dan.ps

Go to the output files for this example

## Command line arguments

       Standard (Mandatory) qualifiers (* if not always prompted):
      [-sequence]          seqall     Sequence database USA
       -windowsize         integer    The values of melting point and other
                                      thermodynamic properties of the sequence are
                                      determined by taking a short length of
                                      sequence known as a window and determining
                                      the properties of the sequence in that
                                      window. The window is incrementally moved
                                      along the sequence with the properties being
                                      calculated at each new position.
       -shiftincrement     integer    This is the amount by which the window is
                                      moved at each increment in order to find the
                                      melting point and other properties along
                                      the sequence.
       -dnaconc            float      Enter DNA concentration (nM)
       -saltconc           float      Enter salt concentration (mM)
    *  -mintemp            float      Enter a minimum value for the temperature
                                      scale (y-axis) of the plot.
    *  -graph              xygraph    Graph type
    *  -outfile            report     If a plot is not being produced then data on
                                      the melting point etc. in each window along
                                      the sequence is output to the file.

       Additional (Optional) qualifiers (* if not always prompted):
       -product            toggle     This prompts for percent formamide, percent
                                      of mismatches allowed and product length.
    *  -formamide          float      This specifies the percent formamide to be
                                      used in calculations (it is ignored unless
                                      -product is used).
    *  -mismatch           float      This specifies the percent mismatch to be
                                      used in calculations (it is ignored unless
                                      -product is used).
    *  -prodlen            integer    This specifies the product length to be used
                                      in calculations (it is ignored unless
                                      -product is used).
       -thermo             toggle     Output the DeltaG, DeltaH and DeltaS values
                                      of the sequence windows to the output data
                                      file.
    *  -temperature        float      If -thermo has been specified then this
                                      specifies the temperature at which to
                                      calculate the DeltaG, DeltaH and DeltaS
                                      values.

       Advanced (Unprompted) qualifiers:
       -rna                boolean    This specifies that the sequence is an RNA
                                      sequence and not a DNA sequence.
       -plot               toggle     If this is not specified then the file of
                                      output data is produced, else a plot of the
                                      melting point along the sequence is
                                      produced.

       Associated qualifiers:

       "-sequence" associated qualifiers
       -sbegin1             integer    Start of each sequence to be used
       -send1               integer    End of each sequence to be used
       -sreverse1           boolean    Reverse (if DNA)
       -sask1               boolean    Ask for begin/end/reverse
       -snucleotide1        boolean    Sequence is nucleotide
       -sprotein1           boolean    Sequence is protein
       -slower1             boolean    Make lower case
       -supper1             boolean    Make upper case
       -sformat1            string     Input sequence format
       -sdbname1            string     Database name
       -sid1                string     Entryname
       -ufo1                string     UFO features
       -fformat1            string     Features format
       -fopenfile1          string     Features file name

       "-graph" associated qualifiers
       -gprompt             boolean    Graph prompting
       -gtitle              string     Graph title
       -gsubtitle           string     Graph subtitle
       -gxtitle             string     Graph x axis title
       -gytitle             string     Graph y axis title
       -goutfile            string     Output file for non interactive displays
       -gdirectory          string     Output directory

       "-outfile" associated qualifiers
       -rformat             string     Report format
       -rname               string     Base file name
       -rextension          string     File name extension
       -rdirectory          string     Output directory
       -raccshow            boolean    Show accession number in the report
       -rdesshow            boolean    Show description in the report
       -rscoreshow          boolean    Show the score in the report
       -rusashow            boolean    Show the full USA in the report

       General qualifiers:
       -auto                boolean    Turn off prompts
       -stdout              boolean    Write standard output
       -filter              boolean    Read standard input, write standard output
       -options             boolean    Prompt for standard and additional values
       -debug               boolean    Write debug output to program.dbg
       -verbose             boolean    Report some/full command line options
       -help                boolean    Report command line options. More
                                      information on associated and general
                                      qualifiers can be found with -help -verbose
       -warning             boolean    Report warnings
       -error               boolean    Report errors
       -fatal               boolean    Report fatal errors
       -die                 boolean    Report deaths

Standard (Mandatory) qualifiers | Allowed values | Default
--- | --- | ---
[-sequence] (Parameter 1) | Sequence database USA | Readable sequence(s) | Required
-windowsize | The values of melting point and other thermodynamic properties of the sequence are determined by taking a short length of sequence known as a window and determining the properties of the sequence in that window. The window is incrementally moved along the sequence with the properties being calculated at each new position. | Integer from 1 to 100 | 20
-shiftincrement | This is the amount by which the window is moved at each increment in order to find the melting point and other properties along the sequence. | Integer 1 or more | 1
-dnaconc | Enter DNA concentration (nM) | Number from 1.000 to 100000.000 | 50.
-saltconc | Enter salt concentration (mM) | Number from 1.000 to 1000.000 | 50.
-mintemp | Enter a minimum value for the temperature scale (y-axis) of the plot. | Number from 0.000 to 150.000 | 55.
-graph | Graph type | EMBOSS has a list of known devices, including postscript, ps, hpgl, hp7470, hp7580, meta, colourps, cps, xwindows, x11, tektronics, tekt, tek4107t, tek, none, null, text, data, xterm, png, xml | EMBOSS_GRAPHICS value, or x11
-outfile | If a plot is not being produced then data on the melting point etc. in each window along the sequence is output to the file. | Report output file |
Additional (Optional) qualifiers | Allowed values | Default
-product | This prompts for percent formamide, percent of mismatches allowed and product length. | Toggle value Yes/No | No
-formamide | This specifies the percent formamide to be used in calculations (it is ignored unless -product is used). | Number from 0.000 to 100.000 | 0.
-mismatch | This specifies the percent mismatch to be used in calculations (it is ignored unless -product is used). | Number from 0.000 to 100.000 | 0.
-prodlen | This specifies the product length to be used in calculations (it is ignored unless -product is used). | Any integer value | Window size (20)
-thermo | Output the DeltaG, DeltaH and DeltaS values of the sequence windows to the output data file. | Toggle value Yes/No | No
-temperature | If -thermo has been specified then this specifies the temperature at which to calculate the DeltaG, DeltaH and DeltaS values. | Number from 0.000 to 100.000 | 25.
Advanced (Unprompted) qualifiers | Allowed values | Default
-rna | This specifies that the sequence is an RNA sequence and not a DNA sequence. | Boolean value Yes/No | No
-plot | If this is not specified then the file of output data is produced, else a plot of the melting point along the sequence is produced. | Toggle value Yes/No | No

## Input file format

Any DNA or RNA sequence USA.

### Input files for usage example

'tembl:paamir' is a sequence entry in the example nucleic acid database 'tembl'

### Database entry: tembl:paamir

    ID   PAAMIR     standard; DNA; PRO; 2167 BP.
    XX
    AC   X13776; M43175;
    XX
    SV   X13776.1
    XX
    DT   19-APR-1989 (Rel. 19, Created)
    DT   17-FEB-1997 (Rel. 50, Last updated, Version 22)
    XX
    DE   Pseudomonas aeruginosa amiC and amiR gene for aliphatic amidase regulation
    XX
    KW   aliphatic amidase regulator; amiC gene; amiR gene.
    XX
    OS   Pseudomonas aeruginosa
    OC   Bacteria; Proteobacteria; gamma subdivision; Pseudomonadaceae; Pseudomonas.
    XX
    RN   [1]
    RP   1167-2167
    RA   Rice P.M.;
    RT   ;
    RL   Submitted (16-DEC-1988) to the EMBL/GenBank/DDBJ databases.
    RL   Rice P.M., EMBL, Postfach 10-2209, Meyerhofstrasse 1, 6900 Heidelberg, FRG.
    XX
    RN   [2]
    RP   1167-2167
    RX   MEDLINE; 89211409.
    RA   Lowe N., Rice P.M., Drew R.E.;
    RT   "Nucleotide sequence of the aliphatic amidase regulator gene of Pseudomonas
    RT   aeruginosa";
    RL   FEBS Lett. 246:39-43(1989).
    XX
    RN   [3]
    RP   1-1292
    RX   MEDLINE; 91317707.
    RA   Wilson S., Drew R.;
    RT   "Cloning and DNA seqence of amiC, a new gene regulating expression of the
    RT   Pseudomonas aeruginosa aliphatic amidase, and purification of the amiC
    RT   product.";
    RL   J. Bacteriol. 173:4914-4921(1991).
    XX
    RN   [4]
    RP   1-2167
    RA   Rice P.M.;
    RT   ;
    RL   Submitted (04-SEP-1991) to the EMBL/GenBank/DDBJ databases.
    RL   Rice P.M., EMBL, Postfach 10-2209, Meyerhofstrasse 1, 6900 Heidelberg, FRG.
    XX
    DR   SWISS-PROT; P10932; AMIR_PSEAE.
    DR   SWISS-PROT; P27017; AMIC_PSEAE.
    DR   SWISS-PROT; Q51417; AMIS_PSEAE.

      [Part of this file has been deleted for brevity]

    FT                   phenotype"
    FT                   /replace=""
    FT                   /gene="amiC"
    FT   misc_feature    1
    FT                   /note="last base of an XhoI site"
    FT   misc_feature    648..653
    FT                   /note="end of 658bp XhoI fragment, deletion in  pSW3 causes
    FT                   constitutive expression of amiE"
    FT   conflict        1281
    FT                   /replace="g"
    FT                   /citation=[3]
    XX
    SQ   Sequence 2167 BP; 363 A; 712 C; 730 G; 362 T; 0 other;
         ggtaccgctg gccgagcatc tgctcgatca ccaccagccg ggcgacggga actgcacgat        60
         ctacctggcg agcctggagc acgagcgggt tcgcttcgta cggcgctgag cgacagtcac       120
         aggagaggaa acggatggga tcgcaccagg agcggccgct gatcggcctg ctgttctccg       180
         aaaccggcgt caccgccgat atcgagcgct cgcacgcgta tggcgcattg ctcgcggtcg       240
         agcaactgaa ccgcgagggc ggcgtcggcg gtcgcccgat cgaaacgctg tcccaggacc       300
         ccggcggcga cccggaccgc tatcggctgt gcgccgagga cttcattcgc aaccgggggg       360
         tacggttcct cgtgggctgc tacatgtcgc acacgcgcaa ggcggtgatg ccggtggtcg       420
         agcgcgccga cgcgctgctc tgctacccga ccccctacga gggcttcgag tattcgccga       480
         acatcgtcta cggcggtccg gcgccgaacc agaacagtgc gccgctggcg gcgtacctga       540
         ttcgccacta cggcgagcgg gtggtgttca tcggctcgga ctacatctat ccgcgggaaa       600
         gcaaccatgt gatgcgccac ctgtatcgcc agcacggcgg cacggtgctc gaggaaatct       660
         acattccgct gtatccctcc gacgacgact tgcagcgcgc cgtcgagcgc atctaccagg       720
         cgcgcgccga cgtggtcttc tccaccgtgg tgggcaccgg caccgccgag ctgtatcgcg       780
         ccatcgcccg tcgctacggc gacggcaggc ggccgccgat cgccagcctg accaccagcg       840
         aggcggaggt ggcgaagatg gagagtgacg tggcagaggg gcaggtggtg gtcgcgcctt       900
         acttctccag catcgatacg cccgccagcc gggccttcgt ccaggcctgc catggtttct       960
         tcccggagaa cgcgaccatc accgcctggg ccgaggcggc ctactggcag accttgttgc      1020
         tcggccgcgc cgcgcaggcc gcaggcaact ggcgggtgga agacgtgcag cggcacctgt      1080
         acgacatcga catcgacgcg ccacaggggc cggtccgggt ggagcgccag aacaaccaca      1140
         gccgcctgtc ttcgcgcatc gcggaaatcg atgcgcgcgg cgtgttccag gtccgctggc      1200
         agtcgcccga accgattcgc cccgaccctt atgtcgtcgt gcataacctc gacgactggt      1260
         ccgccagcat gggcggggga ccgctcccat gagcgccaac tcgctgctcg gcagcctgcg      1320
         cgagttgcag gtgctggtcc tcaacccgcc gggggaggtc agcgacgccc tggtcttgca      1380
         gctgatccgc atcggttgtt cggtgcgcca gtgctggccg ccgccggaag ccttcgacgt      1440
         gccggtggac gtggtcttca ccagcatttt ccagaatggc caccacgacg agatcgctgc      1500
         gctgctcgcc gccgggactc cgcgcactac cctggtggcg ctggtggagt acgaaagccc      1560
         cgcggtgctc tcgcagatca tcgagctgga gtgccacggc gtgatcaccc agccgctcga      1620
         tgcccaccgg gtgctgcctg tgctggtatc ggcgcggcgc atcagcgagg aaatggcgaa      1680
         gctgaagcag aagaccgagc agctccagga ccgcatcgcc ggccaggccc ggatcaacca      1740
         ggccaaggtg ttgctgatgc agcgccatgg ctgggacgag cgcgaggcgc accagcacct      1800
         gtcgcgggaa gcgatgaagc ggcgcgagcc gatcctgaag atcgctcagg agttgctggg      1860
         aaacgagccg tccgcctgag cgatccgggc cgaccagaac aataacaaga ggggtatcgt      1920
         catcatgctg ggactggttc tgctgtacgt tggcgcggtg ctgtttctca atgccgtctg      1980
         gttgctgggc aagatcagcg gtcgggaggt ggcggtgatc aacttcctgg tcggcgtgct      2040
         gagcgcctgc gtcgcgttct acctgatctt ttccgcagca gccgggcagg gctcgctgaa      2100
         ggccggagcg ctgaccctgc tattcgcttt tacctatctg tgggtggccg ccaaccagtt      2160
         cctcgag                                                                2167
    //

## Output file format

If a plot is not being produced, dan reports the sequence of each oligomer window, its melting temperature under the specified conditions and its GC content.

The output is a standard EMBOSS report file.

The results can be output in one of several styles by using the command-line qualifier -rformat xxx, where 'xxx' is replaced by the name of the required format. The available format names are: embl, genbank, gff, pir, swiss, trace, listfile, dbmotif, diffseq, excel, feattable, motif, regions, seqtable, simple, srs, table, tagseq

See: http://emboss.sf.net/docs/themes/ReportFormats.html for further information on report formats.

By default dan writes a 'seqtable' report file.

### Output files for usage example

### File: paamir.dan

    ########################################
    # Program: dan
    # Rundate: Thu Jul 15 2004 12:00:00
    # Report_format: seqtable
    # Report_file: paamir.dan
    ########################################

    #=======================================
    #
    # Sequence: PAAMIR     from: 1   to: 2167
    # HitCount: 2148
    #=======================================

      Start     End     Tm     GC DeltaG DeltaH DeltaS TmProd Sequence
          1      20   64.9   70.0      .      .      .      . ggtaccgctggccgagcatc
          2      21   63.7   65.0      .      .      .      . gtaccgctggccgagcatct
          3      22   63.7   65.0      .      .      .      . taccgctggccgagcatctg
          4      23   66.9   70.0      .      .      .      . accgctggccgagcatctgc
          5      24   66.7   70.0      .      .      .      . ccgctggccgagcatctgct
          6      25   65.5   70.0      .      .      .      . cgctggccgagcatctgctc
          7      26   65.5   70.0      .      .      .      . gctggccgagcatctgctcg
          8      27   63.7   65.0      .      .      .      . ctggccgagcatctgctcga
          9      28   62.9   60.0      .      .      .      . tggccgagcatctgctcgat
         10      29   62.6   65.0      .      .      .      . ggccgagcatctgctcgatc
         11      30   61.7   60.0      .      .      .      . gccgagcatctgctcgatca
         12      31   60.2   60.0      .      .      .      . ccgagcatctgctcgatcac
         13      32   60.2   60.0      .      .      .      . cgagcatctgctcgatcacc
         14      33   59.0   55.0      .      .      .      . gagcatctgctcgatcacca
         15      34   59.2   55.0      .      .      .      . agcatctgctcgatcaccac
         16      35   60.4   60.0      .      .      .      . gcatctgctcgatcaccacc
         17      36   58.9   55.0      .      .      .      . catctgctcgatcaccacca
         18      37   58.6   55.0      .      .      .      . atctgctcgatcaccaccag
         19      38   61.3   60.0      .      .      .      . tctgctcgatcaccaccagc
         20      39   62.4   65.0      .      .      .      . ctgctcgatcaccaccagcc
         21      40   63.9   65.0      .      .      .      . tgctcgatcaccaccagccg
         22      41   64.9   70.0      .      .      .      . gctcgatcaccaccagccgg
         23      42   64.3   70.0      .      .      .      . ctcgatcaccaccagccggg
         24      43   66.1   70.0      .      .      .      . tcgatcaccaccagccgggc
         25      44   67.5   75.0      .      .      .      . cgatcaccaccagccgggcg
         26      45   66.1   70.0      .      .      .      . gatcaccaccagccgggcga
         27      46   66.3   70.0      .      .      .      . atcaccaccagccgggcgac
         28      47   68.6   75.0      .      .      .      . tcaccaccagccgggcgacg
         29      48   69.8   80.0      .      .      .      . caccaccagccgggcgacgg
         30      49   70.7   80.0      .      .      .      . accaccagccgggcgacggg
         31      50   70.5   80.0      .      .      .      . ccaccagccgggcgacggga
         32      51   68.6   75.0      .      .      .      . caccagccgggcgacgggaa
         33      52   68.6   75.0      .      .      .      . accagccgggcgacgggaac
         34      53   68.4   75.0      .      .      .      . ccagccgggcgacgggaact
         35      54   67.4   75.0      .      .      .      . cagccgggcgacgggaactg
         36      55   68.9   75.0      .      .      .      . agccgggcgacgggaactgc

      [Part of this file has been deleted for brevity]

       2101    2120   69.9   80.0      .      .      .      . ggccggagcgctgaccctgc
       2102    2121   68.7   75.0      .      .      .      . gccggagcgctgaccctgct
       2103    2122   65.5   70.0      .      .      .      . ccggagcgctgaccctgcta
       2104    2123   63.5   65.0      .      .      .      . cggagcgctgaccctgctat
       2105    2124   61.3   60.0      .      .      .      . ggagcgctgaccctgctatt
       2106    2125   60.1   60.0      .      .      .      . gagcgctgaccctgctattc
       2107    2126   61.7   60.0      .      .      .      . agcgctgaccctgctattcg
       2108    2127   63.4   65.0      .      .      .      . gcgctgaccctgctattcgc
       2109    2128   61.7   60.0      .      .      .      . cgctgaccctgctattcgct
       2110    2129   59.5   55.0      .      .      .      . gctgaccctgctattcgctt
       2111    2130   57.1   50.0      .      .      .      . ctgaccctgctattcgcttt
       2112    2131   56.4   45.0      .      .      .      . tgaccctgctattcgctttt
       2113    2132   54.7   45.0      .      .      .      . gaccctgctattcgctttta
       2114    2133   55.0   45.0      .      .      .      . accctgctattcgcttttac
       2115    2134   55.9   50.0      .      .      .      . ccctgctattcgcttttacc
       2116    2135   54.7   45.0      .      .      .      . cctgctattcgcttttacct
       2117    2136   52.0   40.0      .      .      .      . ctgctattcgcttttaccta
       2118    2137   51.2   35.0      .      .      .      . tgctattcgcttttacctat
       2119    2138   50.9   40.0      .      .      .      . gctattcgcttttacctatc
       2120    2139   49.0   35.0      .      .      .      . ctattcgcttttacctatct
       2121    2140   49.3   35.0      .      .      .      . tattcgcttttacctatctg
       2122    2141   51.1   35.0      .      .      .      . attcgcttttacctatctgt
       2123    2142   52.2   40.0      .      .      .      . ttcgcttttacctatctgtg
       2124    2143   54.0   45.0      .      .      .      . tcgcttttacctatctgtgg
       2125    2144   55.2   50.0      .      .      .      . cgcttttacctatctgtggg
       2126    2145   53.9   45.0      .      .      .      . gcttttacctatctgtgggt
       2127    2146   52.3   45.0      .      .      .      . cttttacctatctgtgggtg
       2128    2147   53.5   45.0      .      .      .      . ttttacctatctgtgggtgg
       2129    2148   56.0   50.0      .      .      .      . tttacctatctgtgggtggc
       2130    2149   57.8   55.0      .      .      .      . ttacctatctgtgggtggcc
       2131    2150   60.1   60.0      .      .      .      . tacctatctgtgggtggccg
       2132    2151   63.4   65.0      .      .      .      . acctatctgtgggtggccgc
       2133    2152   64.3   70.0      .      .      .      . cctatctgtgggtggccgcc
       2134    2153   63.4   65.0      .      .      .      . ctatctgtgggtggccgcca
       2135    2154   62.7   60.0      .      .      .      . tatctgtgggtggccgccaa
       2136    2155   64.5   65.0      .      .      .      . atctgtgggtggccgccaac
       2137    2156   66.5   70.0      .      .      .      . tctgtgggtggccgccaacc
       2138    2157   66.8   70.0      .      .      .      . ctgtgggtggccgccaacca
       2139    2158   66.8   70.0      .      .      .      . tgtgggtggccgccaaccag
       2140    2159   66.8   70.0      .      .      .      . gtgggtggccgccaaccagt
       2141    2160   65.9   65.0      .      .      .      . tgggtggccgccaaccagtt
       2142    2161   65.6   70.0      .      .      .      . gggtggccgccaaccagttc
       2143    2162   65.6   70.0      .      .      .      . ggtggccgccaaccagttcc
       2144    2163   64.4   65.0      .      .      .      . gtggccgccaaccagttcct
       2145    2164   64.1   65.0      .      .      .      . tggccgccaaccagttcctc
       2146    2165   65.4   70.0      .      .      .      . ggccgccaaccagttcctcg
       2147    2166   64.2   65.0      .      .      .      . gccgccaaccagttcctcga
       2148    2167   62.4   65.0      .      .      .      . ccgccaaccagttcctcgag

    #---------------------------------------
    #---------------------------------------

### Output files for usage example 2

### Graphics File: dan.ps

### File: paamir.dan

The header information contains details of the program, date and sequence

Subsequent lines contain columns of data for each window into the sequence as it is moved along, giving:

- The start postion of the window
- The end position of the window
- The melting temperature of the window
- The percentage C+G of the window
- The sequence of the window

If the qualifier '-product' is used to make the program prompt for percent formamide percent of mismatches allowed and product length, then the output includes the melting temperature of the specified product.

If the qualifier '-thermo' is gived then the DeltaG, DeltaH and DeltaS of the sequence in the window is also output.

## Data files

The EMBOSS data files "Edna.melt" and "Erna.melt" are used to read in the entropy/enthalpy/energy data for DNA and RNA respectively.

EMBOSS data files are distributed with the application and stored in the standard EMBOSS data directory, which is defined by the EMBOSS environment variable EMBOSS_DATA.

To see the available EMBOSS data files, run:

    % embossdata -showall

To fetch one of the data files (for example 'Exxx.dat') into your current directory for you to inspect or modify, run:

    % embossdata -fetch -file Exxx.dat

Users can provide their own data files in their own directories. Project specific files can be put in the current directory, or for tidier directory listings in a subdirectory called ".embossdata". Files for all EMBOSS runs can be put in the user's home directory, or again in a subdirectory called ".embossdata".

The directories are searched in the following order:

- . (your current directory)
- .embossdata (under your current directory)
- ~/ (your home directory)
- ~/.embossdata

## Notes

None.

## References

1. Breslauer, K.J., Frank, R., Blocker, H., and Marky, L.A. (1986). "Predicting DNA Duplex Stability from the Base Sequence." Proceedings of the National Academy of Sciences USA 83, 3746-3750.
2. Baldino, M., Jr. (1989). "High Resolution In Situ Hybridization Histochemistry." In Methods in Enzymology, (P.M. Conn, ed.), 168, 761-777, Academic Press, San Diego, California, USA.

## Warnings

RNA sequences must be submited to this application with the '-rna' qualifier on the command line, otherwise the sequence will be assumed to be DNA.

## Diagnostic Error Messages

None.

## Exit status

0 if successful.

## Known bugs

None.

## See also

Program name | Description
--- | ---
banana | Bending and curvature plot in B-DNA
btwisted | Calculates the twisting in a B-DNA sequence
chaos | Create a chaos game representation plot for a sequence
compseq | Counts the composition of dimer/trimer/etc words in a sequence
freak | Residue/base frequency table or plot
isochore | Plots isochores in large DNA sequences
sirna | Finds siRNA duplexes in mRNA
wordcount | Counts words of a specified size in a DNA sequence

## Author(s)

This program was originally included in EGCG under the names "MELT" and "MELTPLOT", written by Rodrigo Lopez (rls © ebi.ac.uk)
European Bioinformatics Institute, Wellcome Trust Genome Campus, Hinxton, Cambridge CB10 1SD, UK

This application was written by Alan Bleasby (ableasby © rfcgr.mrc.ac.uk)
MRC Rosalind Franklin Centre for Genomics Research Wellcome Trust Genome Campus, Hinxton, Cambridge, CB10 1SB, UK

## History

Written (1999) - Alan Bleasby

## Target users

This program is intended to be used by everyone and everything, from naive users to embedded scripts.

## Comments

None