| fseqbootall

## Wiki

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## Function

Bootstrapped sequences algorithm

## Description

Reads in a data set, and produces multiple data sets from it by bootstrap resampling. Since most programs in the current version of the package allow processing of multiple data sets, this can be used together with the consensus tree program CONSENSE to do bootstrap (or delete-half-jackknife) analyses with most of the methods in this package. This program also allows the Archie/Faith technique of permutation of species within characters. It can also rewrite a data set to convert it from between the PHYLIP Interleaved and Sequential forms, and into a preliminary version of a new XML sequence alignment format which is under development

## Algorithm

SEQBOOT is a general bootstrapping and data set translation tool. It is intended to allow you to generate multiple data sets that are resampled versions of the input data set. Since almost all programs in the package can analyze these multiple data sets, this allows almost anything in this package to be bootstrapped, jackknifed, or permuted. SEQBOOT can handle molecular sequences, binary characters, restriction sites, or gene frequencies. It can also convert data sets between Sequential and Interleaved format, and into the NEXUS format or into a new XML sequence alignment format.

To carry out a bootstrap (or jackknife, or permutation test) with some method in the package, you may need to use three programs. First, you need to run SEQBOOT to take the original data set and produce a large number of bootstrapped or jackknifed data sets (somewhere between 100 and 1000 is usually adequate). Then you need to find the phylogeny estimate for each of these, using the particular method of interest. For example, if you were using DNAPARS you would first run SEQBOOT and make a file with 100 bootstrapped data sets. Then you would give this file the proper name to have it be the input file for DNAPARS. Running DNAPARS with the M (Multiple Data Sets) menu choice and informing it to expect 100 data sets, you would generate a big output file as well as a treefile with the trees from the 100 data sets. This treefile could be renamed so that it would serve as the input for CONSENSE. When CONSENSE is run the majority rule consensus tree will result, showing the outcome of the analysis.

This may sound tedious, but the run of CONSENSE is fast, and that of SEQBOOT is fairly fast, so that it will not actually take any longer than a run of a single bootstrap program with the same original data and the same number of replicates. This is not very hard and allows bootstrapping or jackknifing on many of the methods in this package. The same steps are necessary with all of them. Doing things this way some of the intermediate files (the tree file from the DNAPARS run, for example) can be used to summarize the results of the bootstrap in other ways than the majority rule consensus method does.

If you are using the Distance Matrix programs, you will have to add one extra step to this, calculating distance matrices from each of the replicate data sets, using DNADIST or GENDIST. So (for example) you would run SEQBOOT, then run DNADIST using the output of SEQBOOT as its input, then run (say) NEIGHBOR using the output of DNADIST as its input, and then run CONSENSE using the tree file from NEIGHBOR as its input.

The resampling methods available are:

- The bootstrap. Bootstrapping was invented by Bradley Efron in 1979, and its use in phylogeny estimation was introduced by me (Felsenstein, 1985b; see also Penny and Hendy, 1985). It involves creating a new data set by sampling N characters randomly with replacement, so that the resulting data set has the same size as the original, but some characters have been left out and others are duplicated. The random variation of the results from analyzing these bootstrapped data sets can be shown statistically to be typical of the variation that you would get from collecting new data sets. The method assumes that the characters evolve independently, an assumption that may not be realistic for many kinds of data.
- The partial bootstrap.. This is the bootstrap where fewer than the full number of characters are sampled. The user is asked for the fraction of characters to be sampled. It is primarily useful in carrying out Zharkikh and Li's (1995) Complete And Partial Bootstrap method, and Shimodaira's (2002) AU method, both of which correct the bias of bootstrap P values.
- Block-bootstrapping. One pattern of departure from indeopendence of character evolution is correlation of evolution in adjacent characters. When this is thought to have occurred, we can correct for it by samopling, not individual characters, but blocks of adjacent characters. This is called a block bootstrap and was introduced by Künsch (1989). If the correlations are believed to extend over some number of characters, you choose a block size, B, that is larger than this, and choose N/B blocks of size B. In its implementation here the block bootstrap "wraps around" at the end of the characters (so that if a block starts in the last B-1 characters, it continues by wrapping around to the first character after it reaches the last character). Note also that if you have a DNA sequence data set of an exon of a coding region, you can ensure that equal numbers of first, second, and third coding positions are sampled by using the block bootstrap with B = 3.
- Partial block-bootstrapping. Similar to partial bootstrapping except sampling blocks rather than single characters.
- Delete-half-jackknifing.. This alternative to the bootstrap involves sampling a random half of the characters, and including them in the data but dropping the others. The resulting data sets are half the size of the original, and no characters are duplicated. The random variation from doing this should be very similar to that obtained from the bootstrap. The method is advocated by Wu (1986). It was mentioned by me in my bootstrapping paper (Felsenstein, 1985b), and has been available for many years in this program as an option. Note that, for the present, block-jackknifing is not available, because I cannot figure out how to do it straightforwardly when the block size is not a divisor of the number of characters.
- Delete-fraction jackknifing. Jackknifing is advocated by Farris et. al. (1996) but as deleting a fraction 1/e (1/2.71828). This retains too many characters and will lead to overconfidence in the resulting groups when there are conflicting characters. However it is made available here as an option, with the user asked to supply the fraction of characters that are to be retained.
- Permuting species within characters. This method of resampling (well, OK, it may not be best to call it resampling) was introduced by Archie (1989) and Faith (1990; see also Faith and Cranston, 1991). It involves permuting the columns of the data matrix separately. This produces data matrices that have the same number and kinds of characters but no taxonomic structure. It is used for different purposes than the bootstrap, as it tests not the variation around an estimated tree but the hypothesis that there is no taxonomic structure in the data: if a statistic such as number of steps is significantly smaller in the actual data than it is in replicates that are permuted, then we can argue that there is some taxonomic structure in the data (though perhaps it might be just the presence of aa pair of sibling species).
- Permuting characters. This simply permutes the order of the characters, the same reordering being applied to all species. For many methods of tree inference this will make no difference to the outcome (unless one has rates of evolution correlated among adjacent sites). It is included as a possible step in carrying out a permutation test of homogeneity of characters (such as the Incongruence Length Difference test).
- Permuting characters separately for each species. This is a method introduced by Steel, Lockhart, and Penny (1993) to permute data so as to destroy all phylogenetic structure, while keeping the base composition of each species the same as before. It shuffles the character order separately for each species.
- Rewriting. This is not a resampling or permutation method: it simply rewrites the data set into a different format. That format can be the PHYLIP format. For molecular sequences and discrete morphological character it can also be the NEXUS format. For molecular sequences one other format is available, a new (and nonstandard) XML format of our own devising. When the PHYLIP format is chosen the data set is coverted between Interleaved and Sequential format. If it was read in as Interleaved sequences, it will be written out as Sequential format, and vice versa. The NEXUS format for molecular sequences is always written as interleaved sequences. The XML format is different from (though similar to) a number of other XML sequence alignment formats. An example will be found below. Here is a table to links to those other XML alignment formats:

  Andrew Rambaut's BEAST XML format | http://evolve.zoo.ox.ac.uk/beast/introXML.html and http://evolve.zoo.ox.ac.uk/beast/referenindex.html | A format for alignments. There is also a format for phylogenies described there.
  MSAML M | http://xml.coverpages.org/msaml-desc-dec.html | Defined by Paul Gordon of University of Calgary. See his big list of molecular biology XML projects.
  BSML | http://www.bsml.org/resources/default.asp | Bioinformatic Sequence Markup Language includes a multiple sequence alignment XML format

## Usage

Here is a sample session with fseqbootall

    % fseqbootall -seed 3
    Bootstrapped sequences algorithm
    Input (aligned) sequence set: seqboot.dat
    Phylip seqboot program output file [seqboot.fseqbootall]:

     bootstrap: true
    jackknife: false
     permute: false
     lockhart: false
     ild: false
     justwts: false

    completed replicate number   10
    completed replicate number   20
    completed replicate number   30
    completed replicate number   40
    completed replicate number   50
    completed replicate number   60
    completed replicate number   70
    completed replicate number   80
    completed replicate number   90
    completed replicate number  100

    Output written to file "seqboot.fseqbootall"

    Done.

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

## Command line arguments

    Bootstrapped sequences algorithm
    Version: EMBOSS:6.6.0.0

       Standard (Mandatory) qualifiers:
      [-infilesequences]   seqset     (Aligned) sequence set filename and optional
                                      format, or reference (input USA)
      [-outfile]           outfile    [*.fseqbootall] Phylip seqboot program
                                      output file

       Additional (Optional) qualifiers (* if not always prompted):
       -categories         properties File of input categories
       -mixfile            properties File of mixtures
       -ancfile            properties File of ancestors
       -weights            properties Weights file
       -factorfile         properties Factors file
       -datatype           menu       [s] Choose the datatype (Values: s
                                      (Molecular sequences); m (Discrete
                                      Morphology); r (Restriction Sites); g (Gene
                                      Frequencies))
       -test               menu       [b] Choose test (Values: b (Bootstrap); j
                                      (Jackknife); c (Permute species for each
                                      character); o (Permute character order); s
                                      (Permute within species); r (Rewrite data))
    *  -regular            toggle     [N] Altered sampling fraction
    *  -fracsample         float      [100.0] Samples as percentage of sites
                                      (Number from 0.100 to 100.000)
    *  -rewriteformat      menu       [p] Output format (Values: p (PHYLIP); n
                                      (NEXUS); x (XML))
    *  -seqtype            menu       [d] Output format (Values: d (dna); p
                                      (protein); r (rna))
    *  -morphseqtype       menu       [p] Output format (Values: p (PHYLIP); n
                                      (NEXUS))
    *  -blocksize          integer    [1] Block size for bootstraping (Integer 1
                                      or more)
    *  -reps               integer    [100] How many replicates (Integer 1 or
                                      more)
    *  -justweights        menu       [d] Write out datasets or just weights
                                      (Values: d (Datasets); w (Weights))
    *  -enzymes            boolean    [N] Is the number of enzymes present in
                                      input file
    *  -all                boolean    [N] All alleles present at each locus
    *  -seed               integer    [1] Random number seed between 1 and 32767
                                      (must be odd) (Integer from 1 to 32767)
       -printdata          boolean    [N] Print out the data at start of run
    *  -[no]dotdiff        boolean    [Y] Use dot-differencing
       -[no]progress       boolean    [Y] Print indications of progress of run

       Advanced (Unprompted) qualifiers: (none)
       Associated qualifiers:

       "-infilesequences" 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
       -scircular1         boolean    Sequence is circular
       -squick1            boolean    Read id and sequence only
       -sformat1           string     Input sequence format
       -iquery1            string     Input query fields or ID list
       -ioffset1           integer    Input start position offset
       -sdbname1           string     Database name
       -sid1               string     Entryname
       -ufo1               string     UFO features
       -fformat1           string     Features format
       -fopenfile1         string     Features file name

       "-outfile" associated qualifiers
       -odirectory2        string     Output directory

       General qualifiers:
       -auto               boolean    Turn off prompts
       -stdout             boolean    Write first file to standard output
       -filter             boolean    Read first file from standard input, write
                                      first file to 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 and exit. 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 dying program messages
       -version            boolean    Report version number and exit

Qualifier | Type | Description | Allowed values | Default
--- | --- | --- | --- | ---
Standard (Mandatory) qualifiers
[-infilesequences] (Parameter 1) | seqset | (Aligned) sequence set filename and optional format, or reference (input USA) | Readable set of sequences | Required
[-outfile] (Parameter 2) | outfile | Phylip seqboot program output file | Output file | <*>.fseqbootall
Additional (Optional) qualifiers
-categories | properties | File of input categories | Property value(s) |
-mixfile | properties | File of mixtures | Property value(s) |
-ancfile | properties | File of ancestors | Property value(s) |
-weights | properties | Weights file | Property value(s) |
-factorfile | properties | Factors file | Property value(s) |
-datatype | list | Choose the datatype |
s | (Molecular sequences)
m | (Discrete Morphology)
r | (Restriction Sites)
g | (Gene Frequencies)
| s
-test | list | Choose test |
b | (Bootstrap)
j | (Jackknife)
c | (Permute species for each character)
o | (Permute character order)
s | (Permute within species)
r | (Rewrite data)
| b
-regular | toggle | Altered sampling fraction | Toggle value Yes/No | No
-fracsample | float | Samples as percentage of sites | Number from 0.100 to 100.000 | 100.0
-rewriteformat | list | Output format |
p | (PHYLIP)
n | (NEXUS)
x | (XML)
| p
-seqtype | list | Output format |
d | (dna)
p | (protein)
r | (rna)
| d
-morphseqtype | list | Output format |
p | (PHYLIP)
n | (NEXUS)
| p
-blocksize | integer | Block size for bootstraping | Integer 1 or more | 1
-reps | integer | How many replicates | Integer 1 or more | 100
-justweights | list | Write out datasets or just weights |
d | (Datasets)
w | (Weights)
| d
-enzymes | boolean | Is the number of enzymes present in input file | Boolean value Yes/No | No
-all | boolean | All alleles present at each locus | Boolean value Yes/No | No
-seed | integer | Random number seed between 1 and 32767 (must be odd) | Integer from 1 to 32767 | 1
-printdata | boolean | Print out the data at start of run | Boolean value Yes/No | No
-[no]dotdiff | boolean | Use dot-differencing | Boolean value Yes/No | Yes
-[no]progress | boolean | Print indications of progress of run | Boolean value Yes/No | Yes
Advanced (Unprompted) qualifiers
(none)
Associated qualifiers
"-infilesequences" associated seqset qualifiers
-sbegin1 -sbegin_infilesequences | integer | Start of each sequence to be used | Any integer value | 0
-send1 -send_infilesequences | integer | End of each sequence to be used | Any integer value | 0
-sreverse1 -sreverse_infilesequences | boolean | Reverse (if DNA) | Boolean value Yes/No | N
-sask1 -sask_infilesequences | boolean | Ask for begin/end/reverse | Boolean value Yes/No | N
-snucleotide1 -snucleotide_infilesequences | boolean | Sequence is nucleotide | Boolean value Yes/No | N
-sprotein1 -sprotein_infilesequences | boolean | Sequence is protein | Boolean value Yes/No | N
-slower1 -slower_infilesequences | boolean | Make lower case | Boolean value Yes/No | N
-supper1 -supper_infilesequences | boolean | Make upper case | Boolean value Yes/No | N
-scircular1 -scircular_infilesequences | boolean | Sequence is circular | Boolean value Yes/No | N
-squick1 -squick_infilesequences | boolean | Read id and sequence only | Boolean value Yes/No | N
-sformat1 -sformat_infilesequences | string | Input sequence format | Any string |
-iquery1 -iquery_infilesequences | string | Input query fields or ID list | Any string |
-ioffset1 -ioffset_infilesequences | integer | Input start position offset | Any integer value | 0
-sdbname1 -sdbname_infilesequences | string | Database name | Any string |
-sid1 -sid_infilesequences | string | Entryname | Any string |
-ufo1 -ufo_infilesequences | string | UFO features | Any string |
-fformat1 -fformat_infilesequences | string | Features format | Any string |
-fopenfile1 -fopenfile_infilesequences | string | Features file name | Any string |
"-outfile" associated outfile qualifiers
-odirectory2 -odirectory_outfile | string | Output directory | Any string |
General qualifiers
-auto | boolean | Turn off prompts | Boolean value Yes/No | N
-stdout | boolean | Write first file to standard output | Boolean value Yes/No | N
-filter | boolean | Read first file from standard input, write first file to standard output | Boolean value Yes/No | N
-options | boolean | Prompt for standard and additional values | Boolean value Yes/No | N
-debug | boolean | Write debug output to program.dbg | Boolean value Yes/No | N
-verbose | boolean | Report some/full command line options | Boolean value Yes/No | Y
-help | boolean | Report command line options and exit. More information on associated and general qualifiers can be found with -help -verbose | Boolean value Yes/No | N
-warning | boolean | Report warnings | Boolean value Yes/No | Y
-error | boolean | Report errors | Boolean value Yes/No | Y
-fatal | boolean | Report fatal errors | Boolean value Yes/No | Y
-die | boolean | Report dying program messages | Boolean value Yes/No | Y
-version | boolean | Report version number and exit | Boolean value Yes/No | N

## Input file format

fseqbootall data files read by SEQBOOT are the standard ones for the various kinds of data. For molecular sequences the sequences may be either interleaved or sequential, and similarly for restriction sites. Restriction sites data may either have or not have the third argument, the number of restriction enzymes used. Discrete morphological characters are always assumed to be in sequential format. Gene frequencies data start with the number of species and the number of loci, and then follow that by a line with the number of alleles at each locus. The data for each locus may either have one entry for each allele, or omit one allele at each locus. The details of the formats are given in the main documentation file, and in the documentation files for the groups of programsreads any normal sequence USAs.

### Input files for usage example

### File: seqboot.dat

        5    6
    Alpha     AACAAC
    Beta      AACCCC
    Gamma     ACCAAC
    Delta     CCACCA
    Epsilon   CCAAAC

## Output file format

fseqbootall output will contain the data sets generated by the resampling process. Note that, when Gene Frequencies data is used or when Discrete Morphological characters with the Factors option are used, the number of characters in each data set may vary. It may also vary if there are an odd number of characters or sites and the Delete-Half-Jackknife resampling method is used, for then there will be a 50% chance of choosing (n+1)/2 characters and a 50% chance of choosing (n-1)/2 characters.

The Factors option causes the characters to be resampled together. If (say) three adjacent characters all have the same factors characters, so that they all are understood to be recoding one multistate character, they will be resampled together as a group.

The order of species in the data sets in the output file will vary randomly. This is a precaution to help the programs that analyze these data avoid any result which is sensitive to the input order of species from showing up repeatedly and thus appearing to have evidence in its favor.

The numerical options 1 and 2 in the menu also affect the output file. If 1 is chosen (it is off by default) the program will print the original input data set on the output file before the resampled data sets. I cannot actually see why anyone would want to do this. Option 2 toggles the feature (on by default) that prints out up to 20 times during the resampling process a notification that the program has completed a certain number of data sets. Thus if 100 resampled data sets are being produced, every 5 data sets a line is printed saying which data set has just been completed. This option should be turned off if the program is running in background and silence is desirable. At the end of execution the program will always (whatever the setting of option 2) print a couple of lines saying that output has been written to the output file.

### Output files for usage example

### File: seqboot.fseqbootall

        5     6
    Alpha      AAACCA
    Beta       AAACCC
    Gamma      ACCCCA
    Delta      CCCAAC
    Epsilon    CCCAAA
        5     6
    Alpha      AAACAA
    Beta       AAACCC
    Gamma      ACCCAA
    Delta      CCCACC
    Epsilon    CCCAAA
        5     6
    Alpha      AAAAAC
    Beta       AAACCC
    Gamma      AACAAC
    Delta      CCCCCA
    Epsilon    CCCAAC
        5     6
    Alpha      CCCCCA
    Beta       CCCCCC
    Gamma      CCCCCA
    Delta      AAAAAC
    Epsilon    AAAAAA
        5     6
    Alpha      AAAACC
    Beta       AAACCC
    Gamma      AACACC
    Delta      CCCCAA
    Epsilon    CCCACC
        5     6
    Alpha      AAAACC
    Beta       ACCCCC
    Gamma      AAAACC
    Delta      CCCCAA
    Epsilon    CAAACC
        5     6
    Alpha      AACCAA
    Beta       AACCCC
    Gamma      ACCCAA
    Delta      CCAACC
    Epsilon    CCAAAA
        5     6
    Alpha      AAAACC
    Beta       ACCCCC
    Gamma      AAAACC
    Delta      CCCCAA
    Epsilon    CAAACC
        5     6
    Alpha      AACACC

      [Part of this file has been deleted for brevity]

    Gamma      ACAAAA
    Delta      CCCCCC
    Epsilon    CCAAAA
        5     6
    Alpha      AACAAC
    Beta       AACCCC
    Gamma      AACAAC
    Delta      CCACCA
    Epsilon    CCAAAC
        5     6
    Alpha      AACAAA
    Beta       AACCCC
    Gamma      CCCAAA
    Delta      CCACCC
    Epsilon    CCAAAA
        5     6
    Alpha      ACAAAA
    Beta       ACCCCC
    Gamma      CCAAAA
    Delta      CACCCC
    Epsilon    CAAAAA
        5     6
    Alpha      CAAAAA
    Beta       CCCCCC
    Gamma      CAAAAA
    Delta      ACCCCC
    Epsilon    AAAAAA
        5     6
    Alpha      CAACCC
    Beta       CCCCCC
    Gamma      CAACCC
    Delta      ACCAAA
    Epsilon    AAACCC
        5     6
    Alpha      ACAACC
    Beta       ACCCCC
    Gamma      ACAACC
    Delta      CACCAA
    Epsilon    CAAACC
        5     6
    Alpha      AAAAAA
    Beta       AAAAAC
    Gamma      ACCCCA
    Delta      CCCCCC
    Epsilon    CCCCCA
        5     6
    Alpha      AACAAC
    Beta       AACCCC
    Gamma      CCCAAC
    Delta      CCACCA
    Epsilon    CCAAAC

## Data files

None

## Notes

None.

## References

None.

## Warnings

None.

## Diagnostic Error Messages

None.

## Exit status

It always exits with status 0.

## Known bugs

None.

## See also

Program name | Description
--- | ---
distmat | Create a distance matrix from a multiple sequence alignment
ednacomp | DNA compatibility algorithm
ednadist | Nucleic acid sequence distance matrix program
ednainvar | Nucleic acid sequence invariants method
ednaml | Phylogenies from nucleic acid maximum likelihood
ednamlk | Phylogenies from nucleic acid maximum likelihood with clock
ednapars | DNA parsimony algorithm
ednapenny | Penny algorithm for DNA
eprotdist | Protein distance algorithm
eprotpars | Protein parsimony algorithm
erestml | Restriction site maximum likelihood method
eseqboot | Bootstrapped sequences algorithm
fdiscboot | Bootstrapped discrete sites algorithm
fdnacomp | DNA compatibility algorithm
fdnadist | Nucleic acid sequence distance matrix program
fdnainvar | Nucleic acid sequence invariants method
fdnaml | Estimate nucleotide phylogeny by maximum likelihood
fdnamlk | Estimates nucleotide phylogeny by maximum likelihood
fdnamove | Interactive DNA parsimony
fdnapars | DNA parsimony algorithm
fdnapenny | Penny algorithm for DNA
fdolmove | Interactive Dollo or polymorphism parsimony
ffreqboot | Bootstrapped genetic frequencies algorithm
fproml | Protein phylogeny by maximum likelihood
fpromlk | Protein phylogeny by maximum likelihood
fprotdist | Protein distance algorithm
fprotpars | Protein parsimony algorithm
frestboot | Bootstrapped restriction sites algorithm
frestdist | Calculate distance matrix from restriction sites or fragments
frestml | Restriction site maximum likelihood method
fseqboot | Bootstrapped sequences algorithm

## Author(s)

This program is an EMBOSS conversion of a program written by Joe Felsenstein as part of his PHYLIP package.

Please report all bugs to the EMBOSS bug team (emboss-bug © emboss.open-bio.org) not to the original author.

## History

## Target users

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

## Comments

None