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.

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

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

Contig statistic data of uploaded genome will reviews as shown in previous screen page. click the Go button. 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. and screen page will shown as below: . and genetic code. Several data should be completed about the organism such as taxonomy ID. genus and species name. in this sample Methylomicrobium buryatense.6.

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

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

10. Click the Browse Annotated Genome in SEED view button to obtained information about subsystem feature of the organism. the information is shown in screen page as below: . click the view details button and screen page will seen as below: 11. To see the Job Details. For Methylomicrobium buryatense.

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

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