ena-STUDY-ALFRED WEGENER INSTITUTE FOR POLAR AND MARINE RESEARCH-17-07-2020-14:40:41:175-1519
Plant-derived sedimentary ancient DNA metabarcoding of permafrost core BK-8
In this study we focus on plant-derived sedimentary ancient DNA from the permafrost sediment core BK-8 to track past changes of the terrestrial plant community composition at the Buor Khaya peninsula (Siberia). We used a DNA metabarcoding and amplified the P6-loop of the trnL (UAA) intron (gh primers by Taberlet et al. 2007). In total we processed 54 samples and 5 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 the permafrost sediment core BK-8 to track past changes of the terrestrial plant community composition at the Buor Khaya peninsula (Siberia). We used a DNA metabarcoding and amplified the P6-loop of the trnL (UAA) intron (gh primers by Taberlet et al. 2007). In total we processed 54 samples and 5 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).