The samples for metabarcoding collected during the first campaigns in Cabrera and Islas Cíes have already been processed at the labs in Blanes and Tromso.
We first separated the samples in size fractions using sieves of 10 mm, 1 mm, and 64 microns of mesh size. Then the two larger fractions were combined, resulting in one “large” fraction (> 1 mm) and one “small” fraction (< 1 mm). These fractions were homogenized with a blender. Overall, we had 80 replicate samples. The DNA of these samples was extracted in the lab at Blanes using a commercial kit (Norgen Soil Maxi Kit).
The DNA extracts were sent to the Arctic University of Norway at Tromso, where they were further processed in the laboratory of the team member Owen. The samples were amplified for the two genes chosen, a fragment of the Cytochrome Oxidase I gene, and a fragment of the 12S ribosomal gene, both located in the mitochondria.
It took a concentration of the aliquot of DNA that was kept in Blanes, a second shipment and reamplification of some samples that failed. But finally we managed to obtain the desired amplification products. They were then used to construct DNA libraries and were sent for sequencing to the Novogene firm. We are using a lane of the new high throughput sequencing platform Illumina NovaSeq, and we expect to obtain ca. 50 million sequences.
The samples are on its way to the sequencer right now… we cannot wait to have the results! Thanks to Jesús, Marta, Adrià and Owen for their hard work on the samples.
