Dorolseter-Rondane

The Nedre Dørålseter cabin next to the Rondane National Park in Central Norway. Not a bad place to do field work on a day like this!

In year two of field work for the EBAI-project, we have focused on eDNA and benthic samples at three different stations along the River Atna. By filtering water and analysing DNA from bulk extractions, we want to see how different the diversity of freshwater invertebrates is between stations and if both methods retrieve the same species. As usual, we filtered our water in the field and fixed our 4 min kick samples on ethanol.

Torbjorn-sampling-water-for-DNA

Torbjørn sampling water in sterile PET bottles. Will one liter of river water reveal the diversity of macroinvertebrates living here?

Filtering-water-in-mobile-lab

Markus filtering water in our mobile lab (click here to see a video). Hikers passing were looked suspiciously and probably wondered what kind of dangerous stuff we were doing.

Elisabeth-fixing-benthic-samples

Elisabeth fixing kick-samples of benthic invertebrates. See here for a video of the actual kicksampling.

 

Sorted-samples-EBAIDuring the dark winter months, Markus has been sorting our benthic samples. The deeper we go in the lake, the fewer orders we find, from the left to the right in the figure. Also the abundance of organisms was lower in the samples collected deeper.

 

 

 

 

Sorted samples EBAI 2Diptera were abundant both in the shallow lake benthos and in the river benthos. River sample on the left, lake sample on the right in this figure. Chironomidae were the most abundant Diptera both in the lake and in the river benthos.

 

 

 

 

PCR resultsAfter we extracted the total DNA from the samples, we amplified the barcode section of CO1 gene using different primer sets. We carried out this work in the Hajibabaei lab at the Centre for Biodiversity Genomics, Canada. The amplifications were successful as seen in this figure where we have visualized the amplicons on agarose gel.

 

 

 

 

 

 

Results from Illumina MiSeqThe amplicons were sequenced using Illumina Miseq system in the Hajibabaei lab. The result looks like this on the computer screen. Now we need to collect the good quality sequences, identify them and see whether we can find answers to our questions!

Markus