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Synteny-based annotation software ---------------------------------------------------------- INSTALLATION Step 1 - Install Ruby You can find it here: http://www.ruby-lang.org/en/downloads/ Make sure to include Ruby in your path so that my scripts can find it. Step 2 - Install Rubygems, Trollop, and Blastall You can find these here: http://rubygems.org/pages/download and here: http://ruby.about.com/gi/o.htm?zi=1/XJ&zTi=1&sdn=ruby&cdn=compute&tm=10&f=20&su=p284.12.336.ip_p504.1.336.ip_&tt=2&bt=0&bts=0&zu=http%3A//trollop.rubyforge.org/ You can also install Trollop using rubygems install options. You can find Blastall here: http://www.ncbi.nlm.nih.gov/blast/Blast.cgi?CMD=Web&PAGE_TYPE=BlastDocs&DOC_TYPE=Download Step 3 - Install synteny-based annotator scripts These are all contained in the synteny-based annotator .zip file. Put them all in the directory that you plan to run ruby from. These scripts include the following: synteny_finder.091311.rb fasta_parser.rb blast_pusher.071411.rb blast_job.051909.rb blast_parser.071411.rb synteny_stats.071411.rb synteny-based_annotater.031910.rb annotation_finder.rb synteny_significance_finder.rb Step 4 - Download the most up-to-date prokaryotic genome.faa files from NCBI You can ftp these from here: ftp://ftp.ncbi.nih.gov/genomes/Bacteria/ Even though this directory is called "Bacteria" it also contains the archaeal genomes. The file you want is currently called "all.faa.tar.gz," but someday they may change the name. Put these files in a directory called "ncbi_genomes" Step 5 - Run the "synteny-finder_071411.rb" ruby script on your genome vs. all of the NCBI genome files I would suggest making a ruby script to do this. It might look like this: `ruby synteny_finder.071411.rb -s <output statistics filename> <your genome> <file from ncbi>` You could also use the Dir.foreach method to run my script for each entry in the ncbi genomes' directory. However beware, there are plasmid sequences included in the subdirectory for each ncbi genome. I usually grab the longest .faa file with the Linux wc command to run my script on. This is at least one of the organism's chromosomes instead of a plasmid. You should use one name for the output statistics for all of the ncbi genomes vs. your genome Step 6 - Run the "synteny-based_annotater.031910.rb" script on your genome of interest. Your command line code should look something like this: ruby synteny-based_annotater.031910.rb -q synteny/<query_vs_subject.synteny file> -s synteny/<subject_vs_query.synteny file> -e Bit_score -m list -l synteny_stats_files/<output statistics filename> Output: The output of the synteny statistics file has the following tab-delimited columns: Query gene number, subject gene number, average amino acid ID of orthologs, the average bit score of orthologs, the number of syntenous orthologs, the average length of a syntenous block of genes, and the number of orthologs. The output in the synteny directory has the following tab-delimited columns: Query gene, query gene index, subject gene, subject gene index, percent amino acid identity, length_query_gene/length_subject_gene, average normalized bit score, synteny (0 = no synteny, 1 = synteny), block length in number of genes (nil if the gene is not at the end of a syntenous block), orthology designation (ortholog or possible ortholog), query annotation, subject annotation Known issues: TBA