Report on a training at the Biodiversity Institute of Ontario, University of Guelph

Duration of stay: 04.02. - 23.02.2009

In cooperation of CeDAMar, CAML and the Barcode of Life I was offered a three week stay at the barcoding facilities at the Biodiversity Institute of Ontario at the University of Guelph. The institute hosts the world's leading laboratory for barcoding and is the initiator of BOLD- the Barcode of Life Database. During my three week stay I was given a molecular training in the laboratory facilities, also the barcoding approach, BOLD and opportunities for further collaborations were introduced to me in great detail. The molecular training was carried out on samples imbedded in CeDAMar and CAML projects provided by scientist from all over the world. Also, some of my polychaete samples (190 specimens in total) were processed.

In the beginning of my stay I was shown how samples are generally treated. While for barcoding normally CO I is sequenced, the institute also offers scientists and grad students to work with different markers for different scientific questions. A total of 25 plates (96 wells) with tissue from different invertebrates were available for my training. Due to the limited time of my stay I concentrated on my polychaetes and pycnogonids only. Two different extraction methods were carried out (inverse lysis and CTAB). DNA-extraction is done by a robot that is able to process a 96-well plate in approximately 15 minutes. For PCR standard Folmer-primers are commonly used for invertebrates, different primers can be used or even created if acquired. The whole process from the lysis of tissue samples to the edited sequence takes about 5 days for a 96-well plate. Due to the fact that my polychaetes are rather problematic when it comes to CO I the second period of my stay was characterized by "trouble shooting". Together with several experienced biologists we tried to get the polychaetes to work, unfortunately without any notable results yet. More successful were the pycnogonids which proofed to be an "easy" group for CO I. In the last week of my stay I was introduced to sequence analyses - editing of raw sequences, alignments and building of preliminary trees using BOLD. Also, the opportunities, advantages and disadvantages of BOLD were introduced to me.

As an outcome of this stay I can say that I'm very grateful for the opportunity. Aside from learning about the laboratory facilities and sample procession at the Biodiversity Institute I was able to initiate a collaboration with the institute concerning the barcoding of deep-sea polychaetes and pycnogonids that will also contribute to further CeDAMar projects in the near future.