David Coil deleted Obtain the Full Length 16S Sequence from the Assembly.md  about 9 years ago

Commit id: 699ce9e352092b6dd785b0398c9322f0a63c9010

deletions | additions      

         

#Obtain the Full-Length 16S Sequence from the Assembly    (Skip this step if you are building the tree using the 16S sequence from Sanger sequencing)  1. Go to RAST and sign in  2. On the “Jobs Overview" page, click on view details (under annotation progress) for the microbe you are working with.  3. Click on Browse annotated genome in SEED viewer (At the top of the page)  4. Click on Browse through the features of [organism name]  5. Under Function search for “ssurna” or “SSU rRNA”  (if it doesn't work at first then refresh the page)  6. Find the ssuRNA that is 1400-1800 bp in length (often Illumina assemblies also have fragments of 16S sequence that are only a few hundred bp long)  7. Click on the Feature ID for that sequence  8. Click on the Sequences tab (around the middle of the page )  9. Click on Show Fasta  9. Click on Download Sequences and save as a fasta file. Rename the file to something useful.  11. Double check the identity of the sequence at BLAST:  [http://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome](http://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastn&PAGE_TYPE=BlastSearch&LINK_LOC=blasthome)         

Organism Identification using 16s rRNA gene sequence.md  Library Preparation and Sequencing .md  Genome Assembly and Annotation.md  Obtain the Full Length 16S Sequence from the Assembly.md  Building a 16S rDNA Phylogenetic Tree.md  Data Submission.md  Publication.md