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#1 |
Member
Location: quebec Join Date: Apr 2013
Posts: 35
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Hi All,
I am blasting my miRNA-seq fasta file against rFam data base. Here is the command line I used: blastn -query TOTAL_mapped_reads.fa -task megablast -db Rfam.fasta -out mapped_vs_rfam.txt -outfmt "6 qseqid sseqid qlen evalue pident nident mismatch qcovs qcovhsp" Then I want to retrive those reads which match tRNAs in rFam. There are many hits in the output (see the attached picture). But when I pick the first read which is shown to be 100% match with rRNA in rfam by searching it in rfam website, there is no hit. >62-796337 TCCCATATGGTCTAGCGGTTAGGATTCCTGGTT Briefly, according to my blast result against rfam database, there is a match, but when I search the my sequence in online rfam website, there is no hit. So what is the problem? |
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#2 |
Senior Member
Location: sub-surface moon base Join Date: Apr 2013
Posts: 372
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Based on blastn (at ncbi) your sequence is not a tRNA. The 'best' hit is "PREDICTED: Dasypus novemcinctus WW domain-binding protein 4-like (LOC101430251), transcript variant 2, mRNA".
Perhaps consider a more conservative evalue cutoff? Also, how did you create your Rfam db? "-db Rfam.fasta" looks rather wrong to me but maybe you just named it oddly... |
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#3 | |
Member
Location: quebec Join Date: Apr 2013
Posts: 35
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