Hi Fernan, Thanks for pointing me in the right direction. The problem was indeed with formatdb. But, it wasn't a problem with the version of format db, it was a problem with how it handled a sequence id in my blastable database. I noticed that the erroneous results only occurred for one of the sequences in my blastable database. So I started to investigate that sequence id: The sequence id that lead to erroneous results was: ">8BP_ 5714596" (has underscore and then a space) when I changed it to ">8BP_5714596" (has just underscore) or "> 8BP 5714596" (has just space) the weird results disappeared. So it seems to be a problem with having "_<space>" in the id line in formatdb. I also found that this was also true for 2.2.6, but the discovery happened to coincide with my update so I thought that the blastdb update was the issue. Kind of a weird one! Thanks to everyone for the help. -Bonnie Fernan Aguero <fernan@iib.unsam.edu.ar> To: BHurwitz@twt.com Sent by: cc: biocluster <bioclusters@bioinformatics.org> bioclusters-admin@bioinfo Subject: Re: [Bioclusters] blast 2.2.9 issues with expect value (-e) rmatics.org 06/25/2004 04:15 PM Please respond to bioclusters +----[ BHurwitz@twt.com <BHurwitz@twt.com> (25.Jun.2004 15:56): | | Sequences are appearing to have the same expect value | although the matches in the hsp are very different. The question you should be asking is why the alignments below have the same score. If they have the same score, they'll have the same E value, since the E value is (by definition) the number of alignments with a score >= S (24.3 bits in this case) that you expect to occur by chance. Have you indexed the databases using a 2.2.9 formatdb? Perhaps you're using databases indexed with an older formatdb?. I've also seen strange results myself in the past, and the problem was that somehow the indices created by older versions of formatdb would not be properly read by the newer blast executables. Hope this helps, Fernan | From blast: | | > 5846704 | Length = 177 | | Score = 24.3 bits (12), Expect = 0.16 | Identities = 18/20 (90%) | Strand = Plus / Minus | | Query: 13 ctccagacattgggcgggtt 32 | |||||| ||||| ||||||| | Sbjct: 127 ctccaggcattgagcgggtt 108 | | | > 5714596 | Length = 9401 | | Score = 24.3 bits (12), Expect = 0.16 | Identities = 8/20 (40%) | Strand = Plus / Minus | | | Query: 13 ctccagacattgggcgggtt 32 | | ||| | ||| | Sbjct: 202 tccaagaaaggacccggtcg 183 | | -Bonnie | +----] -- F e r n a n A g u e r o http://genoma.unsam.edu.ar/~fernan _______________________________________________ Bioclusters maillist - Bioclusters@bioinformatics.org https://bioinformatics.org/mailman/listinfo/bioclusters