Salmon Output File Formats

Quantification File

Salmon’s main output is its quantification file. This file is a plain-text, tab-separated file with a single header line (which names all of the columns). This file is named quant.sf and appears at the top-level of Salmon’s output directory. The columns appear in the following order:

Name

Length

EffectiveLength

TPM

NumReads

Each subsequent row describes a single quantification record. The columns have the following interpretation.

  • Name — This is the name of the target transcript provided in the input transcript database (FASTA file).

  • Length — This is the length of the target transcript in nucleotides.

  • EffectiveLength — This is the computed effective length of the target transcript. It takes into account all factors being modeled that will effect the probability of sampling fragments from this transcript, including the fragment length distribution and sequence-specific and gc-fragment bias (if they are being modeled).

  • TPM — This is salmon’s estimate of the relative abundance of this transcript in units of Transcripts Per Million (TPM). TPM is the recommended relative abundance measure to use for downstream analysis.

  • NumReads — This is salmon’s estimate of the number of reads mapping to each transcript that was quantified. It is an “estimate” insofar as it is the expected number of reads that have originated from each transcript given the structure of the uniquely mapping and multi-mapping reads and the relative abundance estimates for each transcript.

Command Information File

In the top-level quantification directory, there will be a file called cmd_info.json. This is a JSON format file that records the main command line parameters with which Salmon was invoked for the run that produced the output in this directory.

Auxiliary Files

The top-level quantification directory will contain an auxiliary directory called aux_info (unless the auxiliary directory name was overridden via the command line). This directory will have a number of files (and subfolders) depending on how salmon was invoked.

Meta information

The auxiliary directory will contain a JSON format file called meta_info.json which contains meta information about the run, including stats such as the number of observed and mapped fragments, details of the bias modeling etc. If Salmon was run with automatic inference of the library type (i.e. --libType A), then one particularly important piece of information contained in this file is the inferred library type. Most of the information recorded in this file should be self-descriptive.

Unique and ambiguous count file

The auxiliary directory also contains 2-column tab-separated file called ambig_info.tsv. This file contains information about the number of uniquely-mapping reads as well as the total number of ambiguously-mapping reads for each transcript. This file is provided mostly for exploratory analysis of the results; it gives some idea of the fraction of each transcript’s estimated abundance that derives from ambiguously-mappable reads.

Observed library format counts

When run in mapping-based mode, the quantification directory will contain a file called lib_format_counts.json. This JSON file reports the number of fragments that had at least one mapping compatible with the designated library format, as well as the number that didn’t. It also records the strand-bias that provides some information about how strand-specific the computed mappings were.

Finally, this file contains a count of the number of mappings that were computed that matched each possible library type. These are counts of mappings, and so a single fragment that maps to the transcriptome in more than one way may contribute to multiple library type counts. Note: This file is currently not generated when Salmon is run in alignment-based mode.

Fragment length distribution

The auxiliary directory will contain a file called fld.gz. This file contains an approximation of the observed fragment length distribution. It is a gzipped, binary file containing integer counts. The number of (signed, 32-bit) integers (with machine-native endianness) is equal to the number of bins in the fragment length distribution (1,001 by default — for fragments ranging in length from 0 to 1,000 nucleotides).

Sequence-specific bias files

If sequence-specific bias modeling was enabled, there will be 4 files in the auxiliary directory named obs5_seq.gz, obs3_seq.gz, exp5_seq.gz, exp5_seq.gz. These encode the parameters of the VLMM that were learned for the 5’ and 3’ fragment ends. Each file is a gzipped, binary file with the same format.

It begins with 3 32-bit signed integers which record the length of the context (window around the read start / end) that is modeled, follwed by the length of the context that is to the left of the read and the length of the context that is to the right of the read.

Next, the file contains 3 arrays of 32-bit signed integers (each of which have a length of equal to the context length recorded above). The first records the order of the VLMM used at each position, the second records the shifts and the widths required to extract each sub-context — these are implementation details.

Next, the file contains a matrix that encodes all VLMM probabilities. This starts with two signed integers of type std::ptrdiff_t. This is a platform-specific type, but on most 64-bit systems should correspond to a 64-bit signed integer. These numbers denote the number of rows (nrow) and columns (ncol) in the array to follow.

Next, the file contains an array of (nrow * ncol) doubles which represent a dense matrix encoding the probabilities of the VLMM. Each row corresponds to a possible preceeding sub-context, and each column corresponds to a position in the sequence context. Unused values (values where the length of the sub-context exceed the order of the model at that position) contain a 0. This array can be re-shaped into a matrix of the appropriate size.

Finally, the file contains the marginalized 0:sup:th-order probabilities (i.e. the probability of each nucleotide at each position in the context). This is stored as a 4-by-context length matrix. As before, this entry begins with two signed integers that give the number of rows and columns, followed by an array of doubles giving the marginal probabilities. The rows are in lexicographic order.

Fragment-GC bias files

If Salmon was run with fragment-GC bias correction enabled, the auxiliary directory will contain two files named expected_gc.gz and observed_gc.gz. These are gzipped binary files containing, respectively, the expected and observed fragment-GC content curves. These files both have the same form. They consist of a 32-bit signed int, dtype which specifies if the values to follow are in logarithmic space or not. Then, the file contains two signed integers of type std::ptrdiff which give the number of rows and columns of the matrix to follow. Finally, there is an array of nrow by ncol doubles. Each row corresponds to a conditional fragment GC distribution, and the number of columns is the number of bins in the learned (or expected) fragment-GC distribution.

Equivalence class file

If salmon was run with the --dumpEq option, then a file called eq_classes.txt will exist in the auxiliary directory. The format of that file is as follows:

N (num transcripts)
M (num equiv classes)
tn_1
tn_2
...
tn_N
eq_1_size t_11 t_12 ... count
eq_2_size t_21 t_22 ... count

That is, the file begins with a line that contains the number of transcripts (say N) then a line that contains the number of equivalence classes (say M). It is then followed by N lines that list the transcript names — the order here is important, because the labels of the equivalence classes are given in terms of the ID’s of the transcripts. The rank of a transcript in this list is the ID with which it will be labeled when it appears in the label of an equivalence class. Finally, the file contains M lines, each of which describes an equivalence class of fragments. The first entry in this line is the number of transcripts in the label of this equivalence class (the number of different transcripts to which fragments in this class map — call this k). The line then contains the k transcript IDs. Finally, the line contains the count of fragments in this equivalence class (how many fragments mapped to these transcripts). The values in each such line are tab separated. Note: The indices for transcripts referenced in this file start at 0.

If salmon was run with the --dumpEqWeights or -d option, then the eq_classes.txt file will include a textual representation of the range-factorized equivalence classes will exist in the auxiliary directory. The format of that file is specified as follows:

N (num transcripts)
M (num equiv classes)
tn_1
tn_2
...
tn_N
eq_1_size t_11 t_12 ... p_11 p_12 ... count
eq_2_size t_21 t_22 ... p_21 p_22 ... count

That is, the file begins with a line that contains the number of transcripts (say N) then a line that contains the number of equivalence classes (say M). It is then followed by N lines that list the transcript names — the order here is important, because the labels of the equivalence classes are given in terms of the ID’s of the transcripts. The rank of a transcript in this list is the ID with which it will be labeled when it appears in the label of an equivalence class. Finally, the file contains M lines, each of which describes a range-factorized equivalence class of fragments. The first entry in this line is the number of transcripts in the label of this equivalence class (the number of different transcripts to which fragments in this class map — call this k). The line then contains the k transcript IDs that partially define the label of this range-factorized equivalence class followed by k floating point values which correspond to the conditional probabilities of drawing a fragment from each of these k transcripts within this range-factorized equivalence class. Finally, the line contains the count of fragments in this equivalence class (how many fragments mapped to these transcripts with approximately this conditional probability distribution). The values in each such line are tab separated. Note: The indices for transcripts referenced in this file start at 0. Note: Unlike the simple equivalence classes, the same transcript set can appear more than once in the set of range-factorized equivalence classes. This is because different sets of fragments can induce quite different conditional probability distributions among these transcripts. For more details on this representation, please check the paper describing range-factorized equivalence classes.