RAXML ML/Thorough Bootstrap raxmlgui makes this command line example: raxmlHPC-PTHREADS.exe -T 2 -b 631 -m PROTGAMMABLOSUM62F -k -p 223 -N 100 -o 1_Euglena_gracilis -s infile2.txt" -n testR -O &&raxmlHPC-PTHREADS.exe -T 2 -f d -m PROTGAMMABLOSUM62F -o 1_Euglena_gracilis -s infile2.txt" -N 1 -n testB -p 730 -O &&raxmlHPC-PTHREADS.exe -T 2 -f b -t RAxML_bestTree.testB -z RAxML_bootstrap.testR -m PROTGAMMABLOSUM62F -s infile2.txt" -n test.tre -O exit NGBW-JOB-RAXMLHPC2_WORKFLOW-42680FE7CE894B5FA2B08B74DDD556B2 (saved) raxmlHPC-HYBRID -s infile.txt -N 100 -m GTRGAMMA -p 12345 -k -O -b 787 -o mycobac_01 -n testS && raxmlHPC-HYBRID -f d -m GTRGAMMA -s infile.txt -N 10 -n testZ -p 333 -O -o mycobac_01 && raxmlHPC-PTHREADS -f b -t RAxML_bestTree.testZ -z RAxML_bootstrap.testS -m GTRGAMMA -s infile.txt -n testout -O -o mycobac_01 ~ RAXMLHPC2_WORKFLOW This test: toolId=RAXMLHPC2_WORKFLOW (required) runtime_=0.50 (default) specify_workflow_=MLTB (no default) datatype_=dna (default=dna, can be others) dna_gtrcat_=GTRGAMMA (no default, a value for the model must be set) invariable_= (default is null) ascertainment_=ASC_ (default is null; ASC_ turns on the option; invariable and asc are mutually exclusive) ascertainment_corr_= (default is lewis, felsenstein an dstamatkis are allowed, the latter two require a file via -q) printbrlength_=1 (print branch lengths, default = 0) outgroup_= (default is no outgroup) disable_seqcheck_=1 (default is 0) mulparambootstrap_seed_val_=12345 (set a seed for the -b bootstrapping option, default 12345) outsuffix_MLTB1_=testR (name the output files from bootstrapping step, default=testR) parsimony_seed_val_MLTB2_=12345 (provide random seed value for MLTB sampling step, default 12345) outsuffix_MLTB2_=testB (name for the output files for -f d ML + thorough bootstrap workflow step 2, default=testB) outsuffix_MLTB3_=outfile (name for the output files of step 3 in the MLTB workflow) we make this command line Commandline is: 'raxmlHPC-HYBRID -n testS -s infile.txt -o 1_Euglena_gracilis -m ASC_GTRGAMMA -N 100 -k -p 12345 -b 787 -O --asc-corr lewis && raxmlHPC-HYBRID -f d -m ASC_GTRGAMMA -s infile.txt -N 10 -n testZ -p 333 -O -o 1_Euglena_gracilis --asc-corr lewis && raxmlHPC-PTHREADS -f b -t RAxML_bestTree.testZ -z RAxML_bootstrap.testS -m ASC_GTRGAMMA -s infile.txt -n testout -O --asc-corr lewis -o 1_Euglena_gracilis' Test passed other model options: If not DNA , set datatype_=(protein/rna/binary/multi) then set this parameter (GTRCAT default; prot_sub_model_=PROTGAMMA/PROTCAT; rna_model_ ;bin_model_=BINCAT/BINGAMMA; multi_model_=MULTICAT/MULTIGAMMA) Then set sub_parameters for protein: prot_matrix_spec_= (DAYHOFF is default) other modifiers can be set: invariable_= - switch - Estimate proportion of invariable sites (GTRGAMMA + I) ascertainment_= switch - ascertainment_corr_= excl - sets the kind of correction used use_emp_freqs_= to use empirical frequencies, protein only outgroup_ - String - Outgroup (one or more comma-separated outgroups, see comment for syntax) upload modifying files constraint_ - InFile - Constraint (-g) binary_backbone_ - InFile - Binary Backbone (-r) partition_ - InFile - Use a mixed/partitioned model? (-q) exclude_file_ - InFile - Create an input file that excludes the range of positions specifed in this file (-E) printbrlength_ - Switch - Print branch lengths (-k)