Datatypes

Galaxy is designed to work with many different datatypes. Upon file upload, datatype can be detected and assigned (when possible) or user specified (before or after load). Datatype is also assigned by tools when output is created. It is important to note that many tools will only accept as input datasets with the appropriate datatype assigned.

A datatype can be altered in two ways:

1. By the appropriate tool to transform the data from one type to another. See tool groups: Convert Formats, SAM Tools, and NGS: QC and manipulation, or perform a search in the top left Tool Search box.
2. With the Edit Attributes form, reached by clicking on the pencil icon inside of a dataset box in the history.

Tool developers: please see admin/datatypes for instructions about adding new datatypes to Galaxy.

Galaxy User Interface: "Format Help" accessed through datatype links on tool forms.

Ab1

A binary sequence file in ab1 format with a '.ab1' file extension. You must manually select this datatype when uploading the file.

Axt

blastz pairwise alignment format. Each alignment block in an '.axt' file contains three lines: a summary line and 2 sequence lines. Blocks are separated from one another by blank lines. The summary line contains chromosomal position and size information about the alignment. It consists of 9 required fields.

BAM

A binary file compressed in the BGZF format with a '.bam' file extension. Also see SAM. http://samtools.sourceforge.net/samtools.shtml

Bed

Tab delimited format (tabular) with a '.bed' file extension. Sometimes the number of fields is noted in the file extension, for example: '.bed3', '.bed4', '.bed6', '.bed12'. Valid BED files contain columns 1-3, 1-4, 1-5, 1-6 or 1-12.

Example BED12: 
chr22 1000 5000 cloneA 960 + 1000 5000 0 2 567,488,0,3512
chr22 2000 6000 cloneB 900 - 2000 6000 0 2 433,399, 0,3601

Coordinates have a "0-based, fully-closed start" and a "0-based, half-open end" and are reported with respect to the forward (+) strand. This is true for any coordinate field in a BED file, including features aligned to the reverse (-) strand. Simply put: the smallest coordinate is used as the "start" field, and the largest coordinate is used as the "end" field (for a sequence aligned to the reverse (-) strand, the "end" would be the beginning of the alignment and the "start" would be the end). This can be confusing when first used. Fortunately, an excellent discussion of the BED coordinate system (also used in Interval and certain other data formats) is at UCSC's genomewiki: http://genomewiki.ucsc.edu/index.php/Coordinate_Transforms

For example, the first 100 bases of a chromosome would be given the coordinates: chromStart=0 and chromEnd=100.

• In a display application, a feature with these coordinates would overlap chromosome bases 1-100. This overlap could be on either the (+) or (-) strand, depending on the strand assignment in column 6.
• In a computational algorithm, a feature with these coordinates would represent a chromosome base range with the mathematical notation of [0,100), representing the 0-based numbered chromosome positions 0-99.

Also, see Interval format for more discussion about ""0-based, half-open start, fully closed end" coordinate system.

BED format does not require a header or custom track and/or browser line, but may have one, starting with a "#" or "track" or "browser" or all three. If extracted from UCSC's Table browser or Downloads area, a BED file may start with a 'bin' column. This is an index number and the column should be removed as a first step prior to any analysis (use tool Text Manipulation: Cut columns). This Cut tool can also be used to rearrange the columns of an Interval file to create a BED file.

3 required fields:

9 optional fields:

The core field definitions above are based on the UCSC Genome Browser BED specification: http://genome.ucsc.edu/FAQ/FAQformat.html#format1

BedGraph

A tabular dataset that is line oriented and represents continuous-valued data. The format was developed at UCSC and the complete specification can be found here: http://genome.ucsc.edu/goldenPath/help/bedgraph.html.

BCF

Binary call format or "binary vcf" format has the extension .bcf. Also see Variant call format or .vcf.

http://vcftools.sourceforge.net/specs.html

http://www.1000genomes.org/wiki/analysis/variant-call-format/bcf-binary-vcf-version-2

Dbn

The Dot-Bracket Notation format (.dbn) is a commonly used format to describe 2D RNA structures, in particular as output for structure predictors that can't predict pseudoknots. Every entry consists of three lines:

1. The header line, to provide information about the sequence (e.g. name, free energy).
2. The sequence line, to provide the sequence of which the structures will be described.

3. The structure (Dot-Bracket) line, describing the structure of the sequence.

   • Two nucleotides that form a bond are indicated with encloding parenthesis and dots represent unbound nucleotides.
   • Pseudoknots are indicated with { .. } or [ .. ].

Example of a Dbn file:

>Sequence1 information - small hairpin (dG=-10.3) ggggaaacccc ((((...))))
>Sequence2 information (dG=0.0)
aaaaaaaaaaa ...........

http://ultrastudio.org/en/Dot-Bracket_Notation

http://rna.urmc.rochester.edu/Text/File_Formats.html#DotBracket

Galaxy Dbn (Dot-Bracket notation) rules:

• The first non-empty line is a header line: no comment lines are allowed.

  • A header line starts with a '>' symbol and continues with 0 or multiple symbols until the line ends.

• The second non-empty line is a sequence line.

  • A sequence line may only include chars that match the Fasta format (https://en.wikipedia.org/wiki/FASTA_format#Sequence_representation) symbols for nucleotides: ACGTURYKMSWBDHVN, and may thus not include whitespaces.
  • A sequence line has no prefix and no suffix.
  • A sequence line is case insensitive.

• The third non-empty line is a structure (Dot-Bracket) line and only describes the 2D structure of the sequence above it.

  • A structure line must consist of the following chars: '.{}[]()'.
  • A structure line must be of the same length as the sequence line, and each char represents the structure of the nucleotide above it.
  • A structure line has no prefix and no suffix.

  • A nucleotide pairs with only 1 or 0 other nucleotides.

    • In a structure line, the number of '(' symbols equals the number of ')' symbols, the number of '[' symbols equals the number of ']' symbols and the number of '{' symbols equals the number of '}' symbols.
• The format accepts multiple entries per file, given that each entry is provided as three lines: the header, sequence and structure line.
• Empty lines are allowed.

Fasta

A sequence in FASTA format consists of a single title line and one or more lines of sequence data wrapped to a consistent length. The first character of the title line is a greater-than (">") symbol.

>sequence1 
atgcgtttgcgtgcatgcgtttgcgtgcatgcgtttgcgtgcatgcgtttgcgtgc
gtcggtttcgttgcatgcgtttgcgtgcatgcgtttgcgtgcatgcgtttgcgtgc
atgcgtttgcgtgc
>sequence2
tttcgtgcgtatagtttcgtgcgtatagtttcgtgcgtatagtttcgtgcgtatag
tttcgtgcgtatagtttcgtgcgtatagtttcgtgcgtatagtttcgtgcgtatag
tggcgcggt

Galaxy FASTA format rules:

• the identifier name for a sequence is the first "word" after the ">" symbol in the title line
• no spaces are between the ">" symbol and the identifier name
• any description content in the title line after the first white space (space, tab, etc.) is ignored by most tools
• identifier names must be unique for each sequence contained within any single FASTA dataset
• all sequence lines are wrapped to the same length (40-80 characters is recommended)

Best practices:

• confirm the format of a FASTA dataset at the start of an analysis project
• if a FASTA dataset is used as a Custom Reference Genome, double check the formatting and fix it as needed. The tool NormalizeFasta can be used in most cases -- see the Custom Reference Genome FAQ for the how-to
• use the same Target Fasta/Custom Genome dataset for all steps to avoid technical processing errors

Troubleshooting:

• Some tools will compensate for inconsistent sequence line wrapping, but others will not. If a fasta dataset has inconsistent wrapping or is not wrapped, use the tool FASTA manipulation -> FASTA Width formatter to reformat.
• Including blank lines in a FASTA dataset will cause errors with many tools. Use the tool Filter and Sort -> Select lines that match an expression with the options "that: NOT Matching" and "the pattern: ^$" to remove.
• Certain sources produce FASTA datasets with non-unique identifier names. Use the tool NGS: QC and manipulation -> Rename sequences to assign unique identifier names.

For more details, please see the Wikipedia FASTA_format entry.

Fastq

While the basic FASTQ format remains somewhat constant, identifiers can change quickly when new technologies are introduced. One of the best resource for understanding current formats is the Wikipedia FASTQ_format entry.

When using Galaxy, many tools require that input FASTQ files be standardized as the first step in an analysis. Use the tool NGS: QC and manipulation -> FASTQ Groomer. Instructions on the tool form explain how to use and the resulting output format.

FastqSolexa

FastqSolexa is the Illumina (Solexa) variant of the Fastq format, which stores sequences and quality scores in a single file:

@seq1
GACAGCTTGGTTTTTAGTGAGTTGTTCCTTTCTTT
+seq1
hhhhhhhhhhhhhhhhhhhhhhhhhhPW@hhhhhh
@seq2
GCAATGACGGCAGCAATAAACTCAACAGGTGCTGG
+seq2
hhhhhhhhhhhhhhYhhahhhhWhAhFhSIJGChO

Or:

@seq1
GAATTGATCAGGACATAGGACAACTGTAGGCACCAT
+seq1
40 40 40 40 35 40 40 40 25 40 40 26 40 9 33 11 40 35 17 40 40 33 40 7 9 15 3 22 15 30 11 17 9 4 9 4
@seq2
GAGTTCTCGTCGCCTGTAGGCACCATCAATCGTATG
+seq2
40 15 40 17 6 36 40 40 40 25 40 9 35 33 40 14 14 18 15 17 19 28 31 4 24 18 27 14 15 18 2 8 12 8 11 9

GFF

GFF lines have nine required fields that must be tab-separated.

• Fields are:
• seqname
• source
• feature
• start (1-based)
• end
• score
• strand
• frame
• group

GFF format is also known as General Feature Format 1 or GFF1. The official specification is at http://www.sanger.ac.uk/resources/software/gff/spec.html (notes in the GFF2 specification describe GFF1).

The UCSC Genome Browser GFF specification: http://genome.ucsc.edu/FAQ/FAQformat.html#format3

GTF

GTF lines have nine required fields that must be tab-separated. (Similar to GFF format)

• Fields are:
• seqname
• source
• feature
• start (1-based)
• end
• score
• strand
• frame
• attribute

GTF format is also known as General Feature Format 2 or GFF2. The official specification is at http://www.sanger.ac.uk/resources/software/gff/spec.html (GFF2 specification).

The UCSC Genome Browser GTF specification: http://genome.ucsc.edu/FAQ/FAQformat.html#format4

TIP When using GTF datasets in Galaxy, the dataset should not contain any header lines or blank lines. Extra comment lines (usually staring with a #) or blank lines at the start and sometimes internal to the file should be removed before using the data.

• Remove the headers (lines that start with a "#") with the Select tool using the option "NOT Matching" with the regular expression: ^#
• Remove blank lines with the Select tool using the option "NOT Matching" with the regular expression: ^$
• Once the formatting is fixed, change the datatype to be gft under Edit Attributes (pencil icon).
• Often gft data will be given the datatype gff by default when formatting problems are present, which works fine with some tools and but not with others. It is a good idea to fix the data first, at the start of an analysis, to avoid confusing tool errors.

GFF3

Similar to GFF and GTF in having nine tab-seperated columns of data at the core of file and having a 1-based start coordinate. However, GFF3 format has data that is grouped differently between lines (and sets of lines), can be hierarchically ordered, and can contain extra content such as FASTA sequence. Seeing the official specification (and online validation tool) for details is highly recommended.

TIP When using GFF3 datasets in Galaxy, the dataset must containly only the single header line and the primary data lines or tools may error. Extra comment lines (###), repeats, and fasta content is not accepted.

When obtaining reference annotation from the Ensembl downloads area, choose the GTF annotation for use with Galaxy's tools. Avoid the GFF3 annotation as it contains this extra content.

Known as General Feature Format 3, or GFF3.

The official specification is at http://www.sequenceontology.org/gff3.shtml.

GMOD also has a nice specification at GFF3_Format.

GFF/GTF/GFF3 datasets are available from many sources and can sometimes be created from other datatypes.

• How to source Human, Mouse, and other common genome GTF reference annnotation data. See Method 6 here.
• The public Main Galaxy instance contains tools to examine and manipulate GFF/GTF files under the tool group Filter and Sort.
• The public Galaxy instance at: http://galaxy.raetschlab.org contains tools to examine GFF files or convert various formats to GFF3 under the tool group GFF Toolkit.
• The Tool Shed contains a repository named fml_gff3togtf that houses a collection of tools to convert various formats into GFF3 format.
• New tools are continually added to the Tool Shed - search with a phase like "GFF" to see if others are available.

Interval

Interval format, also known as "Genomic Intervals" is a datatype developed by the Galaxy team. This format incorporates a 0-based genome coordinates and labels in a tab-delimited file with no requirements on column order. (See BED format for a discussion of the ""0-based, half-open start, fully closed end" coordinate system ). Or, this quick reference:

0-relative positions, half-open start, closed stop: Interval, BED, others [0,100) == 0-100 in files == 0-99 in calculations == 1-100 in display

1-relative positions, closed start and closed stop: MAF, others [1,100] == 1-100 in files == 1-100 in calculations == 1-100 in display

Interval dataset start with definition line that assigns the column attributes. Columns are individually assigned to an Interval attribute CHROM, START, END, NAME, STRAND, SCORE, COMMENT. The columns may be in any order and only CHROM, START, and END are required. To add/assign additional columns, use the "Edit Attributes" form (click on pencil icon in top right corner of dataset).

Most common bioinformatics coordinate data formats can be interpreted as or transformed to Interval format. For example: '.bed' datasets (always), 0-based coordinate '.tab' or '.txt' datasets, 1-based '.GFF/.GTF/.GFF3' datasets (use Convert Formats -> GFF-to-BED), and others.

Example definition line:

#CHROM START END STRAND NAME SCORE COMMENT

Example:
#CHROM START END   STRAND NAME COMMENT chr1   10    100   +      exon myExon
chrX   1000  10050 -      gene myGene

Lav

Lav is the primary output format for BLASTZ. The first line of a '.lav' file begins with '#:lav.'.

MAF

TBA and multiz multiple alignment format. The first line of a .maf file begins with ##maf. This word is followed by white-space-separated "variable=value" pairs. There should be no white space surrounding the =.

SAM

Also see BAM. http://samtools.sourceforge.net/samtools.shtml

Scf

A binary sequence file in 'scf' format with a '.scf' file extension. You must manually select this 'File Format' when uploading the file.

Sff

A binary file in 'Standard Flowgram Format' with a '.sff' file extension.

Tabular (tab delimited)

Any data in tab delimited format (tabular). May end with the extension '.tab' or '.txt'.

VCF

Variant call format has a few different active versions, depending on the source. The first comment line will note which you have. These external links can help with understanding the different .vcf types. It is important to note that many tools will not interpret v4.1 content correctly and it should be avoided as input (as of mid-2013). A compressed version of this data is the Binary call format or "binary vcf" format .bcf.

Current VCF specification:

• https://vcftools.github.io/specs.html

Alternate and Prior:

• http://vcftools.sourceforge.net/specs.html
• http://en.wikipedia.org/wiki/Variant_Call_Format
• http://www.1000genomes.org/wiki/Analysis/vcf4.0
• http://www.1000genomes.org/wiki/Analysis/Variant%20Call%20Format/vcf-variant-call-format-version-41

Wig and bigWig

The wiggle format is 1-based, line-oriented, and ends with the extension '.wig'. Wiggle data is preceded by a required track definition line, which adds a number of options for controlling the default display of this track. The wiggle (WIG) format is for display of dense, continuous data. When large, using the related .bigWig format is recommended.

This information and the .wig and .bigWig formats were developed at UCSC. The official specifications are at:

• http://genome.ucsc.edu/goldenPath/help/wiggle.html
• https://genome.ucsc.edu/goldenPath/help/bigWig.html

Plain text

Any plain text file, commonly ending with the file extension ".txt".