We aim to establish a standard operating procedure for high-throughput DNA-based identification and counting of individual fossil pollen grains from species with known DNA barcodes.
In order to develop the most cost- and time- efficient method, we will test a diverse set of high-throughput single-cell sequencing technologies (HT-SCS). The HT-SCS technologies combine cell isolation techniques, (deoxy)ribonucleic acid-processing methods and next-generation sequencing (NGS) to concurrently analyse the genetic content of hundreds of individual cells. Prior to sequencing, individual cells are physically separated and assayed in reaction chambers down to sub-microliter volumes. Limiting the reaction space to such small volumes has a number of important advantages, including improved reaction efficiency, detection sensitivity at the single-molecule, reduced chances of contamination, and minimized costs of reagents. Therefore, we see these technologies to be perfectly suited for molecularly characterizing and counting of single fossil pollen grains in a time- and cost-effective manner (i.e. ‘single-pollen sequencing’).