DNA of quarantine organisms is scarce. Therefore in this work package an investigation in different protocols to store, transport and multiply DNA samples of these organisms was made. In this work package eight different protocols for long term storage and transport of DNA/RNA samples and WGA products were investigated and tested (e.g. filter, beads, other). GenTegra was chosen as storage medium.
Four kits for whole genome amplification (WGA), a method to multiply DNA, were tested on a subset of organisms from each group (fungi, bacteria, arthropods, nematodes, viruses, phytoplasms). The quality of the individual kits was assessed using different methods: TaqMan PCR, conventional PCR, sequence analysis and gel electrophoresis. Based upon results obtained thus far a WGA kit was selected to be used for the rest of the project.
The samples for ring testing in the work package Validation were prepared and a prototype of DNA bank was established. Several protocols have been evaluated and final protocols have been written. Using these protocols NPPO’s can better handle DNA samples of rare specimen to be used as positive and negative controls in their molecular identification and detection assays.