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. 2021 Aug 10;11:16194. doi: 10.1038/s41598-021-95435-3

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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TLEN calculation. The illustration shows how TLEN is calculated for paired-end reads. The definition from the bam specification says that TLEN or the signed observed template length is the length between the mapped end of the template to the mapped start of the template for all reads mapped to the same template (https://samtools.github.io/hts-specs/SAMv1.pdf). This means the distance between the 5′ end of the leftmost read and the 3′ end of the rightmost read is mapped to the template.