Hi,

we were expecting that both reads of a pair would have different quality scores. Maybe the data below makes our problem a bit more clear.

The majority of pairs have the same quality value for both reads (see below), despite a large difference in the CIGAR string. It is not clear to us, why reads with a cigar string like:
1S1M2I3M1I4M1D2M1D60M1S have the same quality as a perfect match. We would have expected that non-perfect matches have a lesser mapping quality.

  1. Matches which are unique, and do not have any subdominant match:
    251 >= MAPQ >= 255, XT=U
  2. Matches which are unique, and have subdominant matches but a different score:
    175 >= MAPQ >= 181, XT=U
  3. Matches which are putatively unique (not unique, but distinguishable by score):
    119 >= MAPQ >= 127, XT=U
  4. Matches which are a perfect tie:
    78 >= MAPQ >= 90, XT=R.
If I understand this correctly, the cigar string does not contribute to the mapping score?  That would explain it then, though I must admit I feel a bit uncomfortable with reads that were mapped uniquely by splitting them in 10 parts or more (score >= 180).
How do you take care of this? Do you clip the reads manually?

HWI-ST661:153:D0FTJACXX:7:1101:1261:74388       99      chr11   63764389        180     25M108N50M
HWI-ST661:153:D0FTJACXX:7:1101:1261:74388       147     chr11   63764583        180     75M

HWI-ST661:153:D0FTJACXX:7:1101:1261:98650       99      chr4    88128614        255     1S73M1S
HWI-ST661:153:D0FTJACXX:7:1101:1261:98650       147     chr4    88128727        255     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:102235      99      chr5    21883611        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:102235      147     chr5    21883892        180     75M

HWI-ST661:153:D0FTJACXX:7:1101:1261:123302      163     chr5    134262601       180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:123302      83      chr5    134262672       180     1S74M
HWI-ST661:153:D0FTJACXX:7:1101:1261:154720      163     chr6    32827449        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:154720      83      chr6    32827520        180     1S43M3D31M

HWI-ST661:153:D0FTJACXX:7:1101:1261:174904      99      chr17   46973042        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:174904      147     chr17   46973131        180     2S15M1D58M
HWI-ST661:153:D0FTJACXX:7:1101:1261:188083      163     chr22   22764112        255     75M
HWI-ST661:153:D0FTJACXX:7:1101:1261:188083      83      chr22   22764366        255     75M

HWI-ST661:153:D0FTJACXX:7:1101:1261:188990      163     chr7    127361327       180     1S1M2I3M1I4M1D2M1D60M1S
HWI-ST661:153:D0FTJACXX:7:1101:1261:188990      83      chr7    127361431       180     24M86083N51M
HWI-ST661:153:D0FTJACXX:7:1101:1262:7502        99      chr17   40024125        180     33M804N42M
HWI-ST661:153:D0FTJACXX:7:1101:1262:7502        147     chr17   40025749        180     75M

HWI-ST661:153:D0FTJACXX:7:1101:1262:22026       99      chr6    32611046        180     74M1S
HWI-ST661:153:D0FTJACXX:7:1101:1262:22026       147     chr6    32611141        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:72904       163     chr16   33962893        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:72904       83      chr16   33963059        180     75M

HWI-ST661:153:D0FTJACXX:7:1101:1262:120662      99      chr2    107314958       119     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:120662      147     chr2    107315099       119     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:122435      145     chr1    91852782        180     55M
HWI-ST661:153:D0FTJACXX:7:1101:1262:122435      97      chr12   127650913       255     16M1D58M1S

HWI-ST661:153:D0FTJACXX:7:1101:1262:126215      163     chr10   13375888        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:126215      83      chr10   13380697        180     2S73M
HWI-ST661:153:D0FTJACXX:7:1101:1262:136701      163     chr14   50085541        255     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:136701      83      chr14   50085669        255     75M

HWI-ST661:153:D0FTJACXX:7:1101:1262:180551      99      chr5    88047745        180     75M
HWI-ST661:153:D0FTJACXX:7:1101:1262:180551      147     chr5    88057001        180     2M1I3M2I47M

best wishes,
Matthias & Daniela
-- 
Matthias Barann
Institute of Clinical Molecular Biology
Christian Albrechts University Kiel
Schittenhelmstr. 12
D-24105 Kiel, Germany

m.barann@ikmb.uni-kiel.de
+49 - (0)431 - 597 8681 (office)