I have previously written about the range of mapping quality scores (MAPQ) that you might see in BAM/SAM files, as produced by popular read mapping programs. A very quick recap:
- Bowtie 2 generates MAPQ scores between 0–42
- BWA generates MAPQ scores between 0–37
- Neither piece of software describes the range of possible scores in their documentation
- The SAM specification defines the possible ranges of the MAPQ score as 0–255 (though 255 should indicate that mapping quality was not available)
- I advocated that you should always take a look at your mapped sequence data to see what ranges of scores are present before doing anything else with your BAM/SAM files
So what is my latest gripe? Well, I've recently been running TopHat (version 2.0.13) to map some RNA-Seq reads to a genome sequence. TopHat uses Bowtie (or Bowtie 2) as the tool to do the intial mapping of reads to the genome, so you might expect it to generate the same range of MAPQ scores as the standalone version of Bowtie.
But it doesn't.
From my initial testing, it seems that the BAM/SAM output file from TopHat only contains MAPQ scores of 0, 1, 3, or 50. I find this puzzling and incongruous. Why produce only four MAPQ scores (compared to >30 different values that Bowtie 2 can produce), and why change the maximum possible value to 50? I turned to the TopHat manual, but found no explanation regarding MAPQ scores.
Turning to Google, I found this useful Biostars post which suggests that five MAPQ values are possible with TopHat (you can also have a value of 2 which I didn't see in my data), and that these values correspond to the following:
- 0 = maps to 10 or more locations
- 1 = maps to 4-9 locations
- 2 = maps to 3 locations
- 3 = maps to 2 locations
- 50 = unique mapping
The post also reveals that, confusingly, TopHat previously used a value of 255 to indicate uniquely mapped reads. However, I then found another Biostars post which says that a MAPQ score of 2 isn't possible with TopHat, and that the meaning of the scores are as follows:
- 0 = maps to 5 or more locations
- 1 = maps to 3-4 locations
- 3 = maps to 2 locations
- 255 = unique mapping
This post was in reference to an older version of TopHat (1.4.1) which probably explains the use of the 255 score rather than 50. The comments on this post reflect some of the confusion over this topic. Going back to the original Biostars post, I then noticed a recent comment suggesting that MAPQ scores of 24, 28, 41, 42, and 44 are also possible with TopHat (version 2.0.13).
As this situation shows, when there is no official explanation that fully describes how a piece of software should work, it can lead to mass speculation by others. Such speculation can sometimes be inconsistant which can end up making things even more confusing. This is what drives bioinformaticians crazy.
I find it deeply frustrating when so much of this confusion could be removed with better documentation by the people that developed the original software. In this case the documentation needs just one paragraph added; something along the lines of…
Mapping Quality scores (MAPQ)
TopHat outputs MAPQ scores in the BAM/SAM files with possible values 0, 1, 2, or 50. The first three values indicate mappings to 5, 3–4, or 2 locations, whereas a value of 50 represents a unique match. Please note that older versions of TopHat used a value of 255 for unique matches. Further note that standalone versions of Bowtie and Bowie 2 (used by TopHat) produce a different range of MAPQ scores (0–42).
Would that be so hard?