A dot means a match on the forward strand and a comma a match on the reverse 
strand, so there's no difference (aka mismatch) in either read covering that 
position. The last column is indeed Phred score + 33.

FYI, if you were to input multiple BAM files, you'd find the output similar, 
with some of the columns repeated for each of the samples.

Devon

____________________________________________
Devon Ryan, Ph.D.
Email: [email protected]
Tel: +49 (0)178 298-6067
Molecular and Cellular Cognition Lab
German Centre for Neurodegenerative Diseases (DZNE)
Ludwig-Erhard-Allee 2
53175 Bonn, Germany

On Aug 4, 2014, at 10:21 PM, Thiago M. Venancio wrote:

> Hi Devon and TNP,
> 
> Thanks for the feedback. Please allow me one clarification to see if I 
> understood the documentation correctly.
> 
> Thanke the following row from my example:
> 
> supercontig_0   32      C       2       .,      BH
> 
> This means that at supercontig_0 we have a reference C at position 32, with 
> two mapped reads with potential SNPs. However, dot and comman means 
> differences between reads and each strand of the reference sequence. So, if 
> the sequencing method is not strand specific, these SNPs should be excluded. 
> Am I missing something or this point is correct ?
> 
> Finally, how the last column (base quality) should be interpreted ? Should I 
> use the ASCII of the character minus 33 ?
> 
> Sorry if these questions are very basic. I am just trying to make sure I 
> understood the process.
> 
> Best,
> Thiago
> 
> 
> 
> 
> On Mon, Aug 4, 2014 at 5:00 PM, Devon Ryan <[email protected]> wrote:
> Hi Thiagp
> 
> The format, including the last 3 columns is described here: 
> http://samtools.sourceforge.net/pileup.shtml
> 
> Best,
> Devon
> 
> ____________________________________________
> Devon Ryan, Ph.D.
> Email: [email protected]
> Tel: +49 (0)178 298-6067
> Molecular and Cellular Cognition Lab
> German Centre for Neurodegenerative Diseases (DZNE)
> Ludwig-Erhard-Allee 2
> 53175 Bonn, Germany
> 
> On Aug 4, 2014, at 9:48 PM, Thiago M. Venancio wrote:
> 
> > Hi all,
> >
> > I ran the samtools mpileup on a set of mapped reads (in bam format). The 
> > output gave me something like that:
> >
> > supercontig_0   30      T       2       ^g.^g,  BH
> > supercontig_0   31      G       2       .,      AG
> > supercontig_0   32      C       2       .,      BH
> > supercontig_0   1689    T       4       .,^g.^g,        GBA5
> > supercontig_0   1690    A       4       .,.,    GBAD
> > supercontig_0   1691    A       4       .,.,    BAAF
> > supercontig_0   1692    C       4       .$,$.,  EA?C
> >
> > ​I inspected all the documentation I could find over the past few hours and 
> > was unable to find ​a complete explanation for this output. I understand 
> > the first three columns, but not the other three.
> >
> > Can anyone point me the appropriate documentation ?
> >
> > Thanks in advance.
> > Thiago
> > ------------------------------------------------------------------------------
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> 
> 
> 
> -- 
> =================================
> Thiago Motta Venancio, M.Sc., PhD
> http://venancio.openwetware.org/
> =================================


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