METABOLIC RECONSTRUCTION USING THE RAST SERVER

The Rapid Annotation using Subsystem Technology (RAST) Server provides highquality genome annotations for prokaryotes across the whole phylogenetic tree. It makes a
SEED-quality annotation available as a service with a 48 hour turnaround time. The SEED
environment and SEED data structures (most prominently FIGfams) are used to compute the
automatic annotations; however data is not added into the SEED automatically. Once
annotation is completed, genomes can be downloaded in a variety of formats or viewed online.
The genome annotation provided does include a mapping of genes to subsystems and a
metabolic reconstruction.
In this procedure, a mapping of genes to metabolic reconstruction is conducting by the
RAST server using whole genome of Methylomicrobium buryatense (accession number
KB455575.1) as sample of microbial target.
1. Open the RAST server in addresses of http://rast.nmpdr.org/ , and screen page will be seen
as below:

2. Login using your user name and password if you have already have RAST account. If you
don’t have it yet, make your personal account by click on Register a new account button
and follow the instruction from the server.

If you successfully login.3. the screen page of your account will be seen as below: .

click the Use this data and go to step 2 button. After that. and screen page will seen as below: . A prokaryotic genome should be uploaded in either a single FASTA format file or in a Genbank format file. and screen page will be seen as below: 5. Upload the sequence file of genome from your computer by clicking Pilih Berkas button.4. Click the Upload a new genome button to requesting a genome analysis for mapping of metabolite reconstruction.

in this sample Methylomicrobium buryatense. Several data should be completed about the organism such as taxonomy ID. Taxonomy ID can be obtained by click the Look up the taxonomy ID in NCBI button and screen page will seen as below: Enter the name of microbes in Search for area. genus and species name. Contig statistic data of uploaded genome will reviews as shown in previous screen page. and genetic code. and screen page will shown as below: .6. click the Go button.

If the analysis of genome by RAST server is finish. and press the Use the data and go to step 3 button. If it have so. Screen page will seen as below: 7. login to your RAST personal account and screen page of your personal account will seen as below: . and click the Finish the upload button. The screen page will seen as below: 8. in this sample 675511.Obtain the taxonomy ID. Complete the information in the previous screen page to improve RAST server ability to track problems in processing uploaded genome. used it to complete the information about uploaded genome. announcement will sent via email.

9. To see the result of RAST server analysis upon uploaded genome. click the Go to the Jobs Overview button and the Jobs Overview screen page will seen as below: .

For Methylomicrobium buryatense. the information is shown in screen page as below: . click the view details button and screen page will seen as below: 11.10. To see the Job Details. Click the Browse Annotated Genome in SEED view button to obtained information about subsystem feature of the organism.

Explore the Comparative Tools tab.12. an abbreviation for Kyoto Encyclopedia of Genes and Genomes. KEGG. and click the KEGG metabolic analysis button to obtain the KEGG map metabolic pathway for analyzed organism. is a popular data base which used to mapping the metabolic pathway. For Methylomicrobium buryatense. the KEGG map metabolic pathway is shown in screen page as below: .