base modifications tags in SAM file
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19 months ago

Hi everyone !

I'm currently working on parsing SAM file to extract methyllation site from a nanopore sequencing. According to the SAM documentation, it's possible thanks to MM:Z and ML:B:C tags.

You can find bellow, a read, the MM:Z and ML:B:C tags extracted from my SAM file:

TGATCGCGCGGACCTGTTCTACCAGGTAGGTCACCGGGTCAAATGATATTTTGATGGTGTTGGACACCACCGTCTGGCTGGCGCTCAGGGTGCCGGAGTTCAGAGCGTAGATGAATGTCTCAAACGCGGAGGATTTCTCGCCTCCCAGCATGTAAATTGGCCACTGCAGGGCGCTGCTCTTGTCAGTATAGCGGAAATGTATGGGGAGCGGCATATTTCGTTAAGGACGGTTGCAATGGCTACCCCAGAATCTTGGCTGCTGTTGCCTTCGACCGCCGCGTTCACGCGCTCAATTGTGGGGTGGAGCACAGCGATCGCTGAAGCGGCGCACGAAAAACGCGAAAGCGTTTCACGATAAATGCGAAAACCACAAAGACACCGACAACTTTCTTCGAGCAAATTCACCTACGCCAGCAACTGAACGAAGTAC

MM:Z:C+m?,1,2,1,0,1,1,0,1,0,0,0,1,2,0,9,0,0,0,0,0,10,2,1,7,4,8,3,0,3,4,4,4,9,6,0,0;

ML:B:C,234,246,49,84,5,0,11,228,254,0,0,2,8,1,3,1,1,0,146,0,10,0,1,115,19,0,167,19,0,121,21,9,188,112,93,6

The sequence has a length of 430 bases. According to the MM:Z tag, it gives information about 5-Methylcytosine presence. The modification status of the first cytosine is unknow, the second is called, the third and the fourth are not called etc... So MM:Z tag gives information for SUM(numeric value of MM:Z) + nb of numeric value = 120 cytosines. ML:B:C tag gives the same number of information about cytosines. However, my read contains only 110 cytosines. Why is it possible to get this difference ?

Best regards.

Antoine

methylation SAM tag nanopore bam • 2.1k views
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Hi, did you find an answer for this question? I have the exact same problem and just don't understand why there are more Cs than indicated.

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Can you share your bam file and point to me which records have such issue?

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19 months ago

There is software to do this for you - modkit this is the modern toolkit from Nanopore, I haven't tried it yet, but think it's intended for bams produced and annotated by dorado.

https://github.com/nanoporetech/modkit

Another older version was called modbamtobed, which I used successfully for modified bams from megalodon.

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