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Michael V. Westbury

Publications and source records attributed to Michael V. Westbury.

2 recordsLinked to original sources

PlainMap: a lightweight, restartable mapping pipeline for ancient and modern DNA

Mapping sequencing reads to a reference genome requires preprocessing and alignment choices that can vary with library type, fragment length, sequencing platform, and reference genome. These considerations are particularly important for ancient and historical DNA, where short and damaged fragments can make the most appropriate mapping strategy difficult to determine a priori. We present PlainMap, a lightweight and restartable mapping pipeline for modern and degraded DNA sequencing data. PlainMap accepts a simple manifest of FASTQ files, automatically identifies single-end and paired-end data from read headers, supports mixed sequencing platforms, and provides alternative mapping strategies for modern and degraded DNA. Deterministic chunking and checkpoint-based execution allow large analyses to resume after interruption, while optional pilot subsampling enables empirical comparison of mapping strategies using identical subsets of raw fragments. PlainMap produces duplicate-filtered BAM files together with fragment-aware mapping and coverage statistics. Evaluation using heterogeneous sequencing data confirmed the expected behaviour of the three mapping modes, while adaptive chunking reduced peak memory use by approximately 25% and allowed an interrupted analysis to resume from completed mapping chunks. PlainMap is implemented as a single Bash script and is freely available at https://github.com/BiodiversityExtinction/PlainMap.

q-bio.GN↗

Sequencing single-stranded libraries on the Illumina NextSeq 500 platform

Single-stranded libraries generated from ancient DNA extracts have specific sequencing requirements. The short library molecules typically associated with ancient DNA templates are expected to behave differently during the annealing to the flow cell and subsequent cluster generation (bridge PCR), requiring optimisation of loading amounts to obtain optimal and consistent cluster densities. For the past 3 years, we have carried out sequencing of single-stranded libraries on the Illumina NextSeq 500 sequencing platform at the Institute for Biochemistry and Biology, University of Potsdam. We report our optimisations here. This document may be useful for other researchers wishing to sequence single-stranded libraries on the NextSeq 500 platform. It does not replace the excellent documentation provided by Illumina (links provided below), but rather serves as additional information specific to single-stranded libraries. The original papers describing the library protocol should also be studied in detail, and complement the information presented here.

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