AlignFASTAinput,writealignedFASTA(AFA)output:
muscle -align input.fa -output aln.afa
Align large input using Super5 algorithm if -align is too expensive, typically needed with more than a
few hundred sequences:
muscle -super5 input.fa -output aln.afa
Singlereplicatealignment:
muscle -align input.fa -perm PERM -perturb SEED -output aln.afa muscle -super5 input.fa -perm PERM
-perturb SEED -output aln.afa
PERM is guide tree permutation none, abc, acb, bca (default none). SEED is perturbation seed 0,
1, 2... (default 0 = don't perturb).
Ensemble of replicate alignments, output in Ensemble FASTA (EFA) format, EFA has one aligned FASTA for
each replicate with header line "<PERM.SEED":
muscle -align input.fa -stratified-output stratified_ensemble.efa muscle -align input.fa
-diversified-output diversified_ensemble.afa
-replicates N
Number of replicates, defaults 4, 100, 100 for stratified,
diversified, resampled. With -stratified there is one replicate per guide tree permutation, total
is 4 x N.
Generate resampled ensemble from existing ensemble by sampling columns with replacement:
muscle -resample ensemble.efa -output resampled.efa
-maxgapfract F
Maximum fraction of gaps in a column (F=0..1, default 0.5).
-minconf CC
Minimum column confidence (CC=0..1, default 0.5).
If ensemble output filename has @, then one FASTA file is generated for each replicate where @ is
replaced by perm.s, otherwise all replicates are written to one EFA file.
Calculatedispersonofanensemble:
muscle -disperse ensemble.efa
ExtractreplicatewithhighesttotalCC(diversifiedinputrecommended):
muscle -maxcc ensemble.efa -output maxcc.afa
ExtractalignedFASTAfilesfromEFAfile:
muscle -efa_explode ensemble.efa
ConvertFASTAtoEFA,inputhasonefilenameperline:
muscle -fa2efa filenames.txt -output ensemble.efa
Update ensemble by adding two sequences of digits to each replicate, digits are column confidence (CC)
values, e.g. "73" means CC=0.73, "++" is CC=1.0:
muscle -addconfseqs ensemble.efa -output ensemble_cc.efa
Calculate letter confidence (LC) values, -ref specifies the alignment to compare against the ensemble
(e.g. from -maxcc), output is in aligned FASTA format with LC values 0, 1 ... 9 instead of letters:
muscle -letterconf ensemble.efa -ref aln.afa -output letterconf.afa
-html aln.html
Alignment colored by LC in HTML format.
-jalview aln.features
Jalview feature file with LC values and colors.
Moredocumentationat:
https://drive5.com/muscle