Getting Started with trimAl v1.2

Thank you for choosing trimAl v1.2 to trim your alignments. In this version we have solved some bugs from the previous version and we have also added new functionality to the program. You will see that it is very easy to get familiar with the program. The first thing you need to do to start is to decide whether you will be using trimAl in its command line version or through the web interface. The command line version is faster and has more possibilities, so it is recommended if you are going to use trimAl extensively.

The trimAl webserver included in Phylemon 2.0 provides a friendly user interface and the opportunity to concatenate your trimmed alignment to many different phylogenetic analysis.

Input and Output formats

trimAl reads and renders protein or nucleotide alignments in several Multiple Sequence Alignment formats, including Phylip, Fasta, Clustal, NBRF/Pir, Mega and Nexus. The program detects automatically the input format and generates the output file in the same format. Alternatively, the user can select a different format for the output. Moreover, trimAl can provide as an output the complementary MSA, that is, the columns that would otherwise be removed by the specified parameters (option -complementary). Finally, to facilite the visualization of trimAl's trimming, the program can generate an html file in which selected and trimmed columns are colored differently (-htmlout).

Besides MSAs, trimAl can optionally produce other outputs, which have been deemed of interest. For instance, to facilitate the tracking of the correspondences between the columns in the original and the trimmed alignment, trimAl can return the relationship between their column numbers (option -colnumbering). trimAl can provide information on gap and/or conservation scores in a MSA. This information can be relative to each column or the distribution of these values along the alignment (options -sgc for gaps and -scc for conservation values) (options -sgt and -sct for gaps and conservation distribution, respectively). When comparing several alignments, trimAl can also offer statistical information about their consistency score (options -sfc for each column and -sft for whole alignment). Finally, trimAl can provide a comparison matrix summarizing the percentage of identities between each pair of sequences in the alignment, their averages and the highest identity pair for each sequence (option -sident).

Command line version

You would need first to install trimAl v1.2 on your computer. On the following link downloads you have the necessary files and instructions to get trimAl properly installed on your computer, whether you are a Linux, MacOS or Windows user.

Once you have trimAl v1.2 installed, just type “trimal” on your prompt to get the basic commands you can use.


In this section, we are going to use one (dataset/example1.phy) of the files included into the trimAl package. We also are going to use the trimAl -htmlout option to show you the trimAl's performance.

A very common way of using trimAl v1.2 to trim an alignment is to use just a gap threshold (the minimum fraction of sequences without a gap that you require to consider a column of “enough quality”)

For example:


   trimal -in example1 -out output1 -htmlout output1.html -gt 1

will remove all columns with any gap (equivalent to -nogaps option)

   trimal -in example1 -out output2 -htmlout output2.html -gt 0.8 -st 0.001

will remove all columns with gaps in more than 20% of the sequences or with a similarity score lower than 0.001

If you feel that, for some alignments this will be too strict and prefer to use a minimum coverage in the trimmed alignment (that is, the trimmed alignment will retain a given percentage of the columns in the original alignment) you can do it as follows:

   trimal -in example1 -out output3 -htmlout output3.html -gt 0.8 -st 0.001 -cons 60

will remove all columns with gaps in more than 20% of the sequences or with a similarity score lower than 0.001, unless this removes more than 40% of the columns in the original alignment, we want to conserve at least 60% of them. In such cases trimAl v1.2 will add the necessary number of columns (in decreasing order of scores) so that the minimum coverage is respected.

Yet another threshold that you can use is based on the comparison of different alignments. Sometimes one does not know which alignment algorithm will perform best (or which parameters, e.g gap penalties). A way out is to just produce different alignments with the different algorithms and then choose the alignment that contains the most consistent residue-pairings, that is the residue pairs that are recovered by most algorithm.


trimAl v1.2 can do this for you, just provide in the input file a list of the paths for the different alignments (in this case, we are going to use the file dataset/fileset1). Just type

   trimal -compareset fileset1 -out output4

You can then trim the output alignment, the most consistency one, with other algorithms or/and trim it based on the consistency values, for instance:

   trimal -compareset fileset1 -out output5 -htmlout output5.html -ct 0.5

Will trim such the most consistency alignment removing all columns with a consistency score lower than 0.5

   trimal -compareset fileset1 -out output5 -ct 0.5 -gt 0.75 -st 0.001 

Will trim such the most consistency alignment removing all columns with a consistency score lower than 0.5, with gaps in more than 25% of the sequences and with a similarity score lower than 0.001. In this case, it is impossible to generate an html file in order to track the trimAl's trimming because we are applying two consecutive trimming methods. The first one is related to the consistency score and the second one is related to the gaps and similarity scores.


Moreover, you can use one of the implemented methods to set up, depending on the alignment features, the different thresholds. Among these methods, you can find -gappyout, -strict and -strictplus as automated methods that uses gaps and similarities distribution to fix the thresholds. You also can find the -nogaps and -noallgaps methods that let you remove columns with, at least, one gap and columns with only gaps respectively. Finally, you can find the heuristic method -automated1 that it is used to decide which is the best automated method to trim your input alignment depending on its features. If you want to know about these methods, please, see our publication for more details.

   trimal -in example1 -out output6 -htmlout output6.html -gappyout

will remove columns from the input alignment using the gappyout method.

   trimal -in example1 -out output7 -htmlout output7.html -strict

will remove columns from the input alignment using the strict method.

   trimal -in example1 -out output8 -htmlout output8.html -automated1

will remove columns from the input alignment using the heuristic automated1 method to decide which is the best automated method to trim the alignment between gappyout and strict ones.


Finally, trimAl can remove spurious sequences from your alignment. For that purpose, it is important to define the -resoverlap and -seqoverlap properly.

   trimal -in example1 -out output9 -htmlout output9.html -resoverlap 0.75 -seqoverlap 80

will remove all those sequences that not earn, at least, to the 80% of residues that achieve an overlap, with the rest of the sequences, of 0.75.

Webserver version

Alternatively you can use trimAl v1.2 through its user-friendly web interface, implemented in Phylemon, an online platform for phylogenetic and evolutionary analyses of molecular sequence data.



Instructions on how to use it can be found here.

getting_started_with_trimal_v1.2.txt · Last modified: 2009/07/21 11:01 by scapella
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