Submitting a Query Sequence


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Point your browser to www.ncbi.nih.gov. Click on the “BLAST” button.
On the BLAST page, click on “BLASTP”, as shown in the thumbnail to the left.


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Previously, a file with the sequence of the HFE protein in FASTA format was saved to the desktop. Open this file, copy the FASTA file (including identifier, description line and sequence) and paste into the BLASTP search window as shown.


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The default setting is to search for matches in all organisms. Often this is unnecessary, and only lengthens the time to complete the search. A BLAST search can be made more efficient by using the pull-down menu - as shown in the graphic to the left - to limit alignments to a single taxon.

If the pull-down menu does not contain the limiting term which is desired, the scientific name, with the [ORGN] qualifier, can be typed into the "Limit by entrez Query" box  (example: Canis familiaris [ORGN]).

In the “Options” box, limit the BLAST search by using the pull-down menu to select “Mammalia”. This will limit the “hits” to only similar sequences which occur in Mammals.

Leave the other options at their default values.
 



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Click on the  button below the Search boxes or at the bottom of the page to submit your query. The screen shown in the graphic to the left should appear, which indicates that the search has been submitted successfully to the BLAST server at NCBI. 

Although the search has been placed into a queue, it will only take approximately 8 seconds to search thousands of sequences and millions of amino acids! 


A conserved domain is a recurring sequence pattern or motif which occurs within a larger sequence. A conserved domain is associated with a characteristic structure and function, and in concert with other domains, determines the function of the entire protein.
 

Click on the  button to open a new browser window that contains the BLAST results. 
 

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