Sequencing Library Preparation from Degraded Samples for Non-illumina Sequencing Platforms

Methods Mol Biol. 2019:1963:85-92. doi: 10.1007/978-1-4939-9176-1_10.

Abstract

Efficient methods for building genomic sequencing libraries from degraded DNA have been in place for Illumina sequencing platforms for some years now, but such methods are still lacking for other sequencing platforms. Here, we provide a protocol for building genomic libraries from degraded DNA (archival or ancient sample material) for sequencing on the Ion Torrent™ high-throughput sequencing platforms. In addition to a reduction in time and cost in comparison to commercial kits, this protocol removes purification steps prior to library amplification, an important consideration for work involving historical samples. Libraries prepared using this method are appropriate for either shotgun sequencing or enrichment-based downstream approaches.

Keywords: Archival DNA; Degraded samples; Ion Torrent™ PGM; Non-illumina platforms; Sequencing libraries.

MeSH terms

  • DNA Damage*
  • DNA, Ancient / analysis*
  • DNA, Ancient / chemistry
  • DNA, Ancient / isolation & purification
  • Gene Library*
  • High-Throughput Nucleotide Sequencing / methods*
  • Sequence Analysis, DNA / methods*

Substances

  • DNA, Ancient