This is the annotation package for insect mitochondrial gene annotation. To run the pipeline, just type: perl MT_annotation_BGI.pl There is no need to compile the scipt. Two perl package is needed: FindBin and File::Basename. There is a test dataset of Drosophila melanogaster in the folder test_data for a quick run. You could follow the command line in the shell scitp: sample_command_line.sh. perl ../MT_annotation_BGI.pl Drosophila_melanogaster_MT.fas ../MT_database/MitoProtein_RefSeq_110502.fasta ./ Then you will get a bunch of files: # The original scaffold file and the built BLAST library: Drosophila_melanogaster_MT.fas Drosophila_melanogaster_MT.fas.nhr Drosophila_melanogaster_MT.fas.nin Drosophila_melanogaster_MT.fas.nsq formatdb.log # the script for tBLASTn MitoProtein_RefSeq_110502.fasta.tblastn.shell # the results of tBLASTn alignment MitoProtein_RefSeq_110502.fasta.blast # filtered results of tBLASTn MitoProtein_RefSeq_110502.fasta.blast.filter # the script for solar MitoProtein_RefSeq_110502.fasta.solar.shell # reformated tBLASTn results MitoProtein_RefSeq_110502.fasta.blast.solar # the filtered solar results MitoProtein_RefSeq_110502.fasta.blast.solar.filter # reformated solar results for clustering MitoProtein_RefSeq_110502.fasta.blast.solar.filter.table # non-redundant solar files MitoProtein_RefSeq_110502.fasta.blast.solar.filter.table.nonredundance # the corresponding information of the non-redundant hits MitoProtein_RefSeq_110502.fasta.blast.solar.filter.nr # the scripts for genewise annotation MitoProtein_RefSeq_110502.fasta.genewise.shell # the genewise results MitoProtein_RefSeq_110502.fasta.genewise # length of each sequences in references MitoProtein_RefSeq_110502.fasta.length # gff format of annotation results MitoProtein_RefSeq_110502.fasta.solar.genewise.gff # the corresponding nucleotide and protein sequences of annotated genes, the final annotated genes MitoProtein_RefSeq_110502.fasta.solar.genewise.gff.cds MitoProtein_RefSeq_110502.fasta.solar.shell The mitochondrial database is the RefSeq MT proteins of different genes. You could update this database and just change the database file in the command line. The folder "blast" and "solar" contains the software of BLAST and SOLAR. You could also download new versions and replace the old one. Please keep in mind that the new version of software must have the same name as old ones.