ena-STUDY-ALFRED WEGENER INSTITUTE FOR POLAR AND MARINE RESEARCH-17-07-2020-14:02:10:526-1515
Plant-derived sedimentary ancient DNA metabarcoding of permafrost cores L14
In this study we focus on plant-derived sedimentary ancient DNA from permafrost sediments to track past changes of the terrestrial plant community composition at Bol'shoy Lyakhovsky Island from MIS 5 to MIS 1. We used a DNA metabarcoding and amplified the P6-loop of the trnL (UAA) intron (gh primers by Taberlet et al. 2007). The samples were collected from frozen permafrost sediment cores L14-02, L14-03, L14-04 (including some larger samples called hand-pieces L14-04B and L14-04C that were taken out manually from the coastal bluff) and L14-05. In total we processed 72 samples (~2 samples per m depth) and 8 extraction blanks. PCRs were performed with dual-tagged primers. For each sample two positive PCR products were pooled and purified under the condition that the extraction and PCR negative controls remained negative. Subsequently, all purified PCR products (comprising samples, extraction blanks and and PCR no template controls (NTCs)) were pooled in equal concentrations for parallel high-throughput sequencing. The library was prepared by Fasteris SA sequencing service following the MetaFast protocol with the HiSeq SBS V4 kit. The library was sequenced on the Economy Lane on the Illumina HiSeq 2500 instrument (2 × 125 bp).
In this study we focus on plant-derived sedimentary ancient DNA from permafrost sediments to track past changes of the terrestrial plant community composition at Bol'shoy Lyakhovsky Island from MIS 5 to MIS 1. We used a DNA metabarcoding and amplified the P6-loop of the trnL (UAA) intron (gh primers by Taberlet et al. 2007). The samples were collected from frozen permafrost sediment cores L14-02, L14-03, L14-04 (including some larger samples called hand-pieces L14-04B and L14-04C that were taken out manually from the coastal bluff) and L14-05. In total we processed 72 samples (~2 samples per m depth) and 8 extraction blanks. PCRs were performed with dual-tagged primers. For each sample two positive PCR products were pooled and purified under the condition that the extraction and PCR negative controls remained negative. Subsequently, all purified PCR products (comprising samples, extraction blanks and and PCR no template controls (NTCs)) were pooled in equal concentrations for parallel high-throughput sequencing. The library was prepared by Fasteris SA sequencing service following the MetaFast protocol with the HiSeq SBS V4 kit. The library was sequenced on the Economy Lane on the Illumina HiSeq 2500 instrument (2 × 125 bp).