Comparing the efficiency of six common methods for DNA extraction from root-lesion nematodes (Pratylenchus spp.)

Orlando, V., Edwards, S.G., Neilson, R., Prior, T., Roberts, D. and Back, M.A. (2020) Comparing the efficiency of six common methods for DNA extraction from root-lesion nematodes (Pratylenchus spp.). Nematology.

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Abstract

Robust and accurate identification of root-lesion nematodes (Pratylenchus spp.) is an essential step for determining their potential threat to crop yields and, consequently, development of an efficient agronomic management strategy. It is recognised that DNA-based techniques provide rapid identification of a range of plant-parasitic nematodes including Pratylenchus spp. Efficient and repeatable DNA extraction is central to molecular methodologies. Here, six common DNA extraction protocols were compared to evaluate their efficiency to obtain quality DNA samples for Pratylenchus penetrans. Samples with five and ten individuals of P. penetrans were successfully extracted and amplified by all extraction methods tested, whereas samples with a single nematode presented challenges for DNA amplification. Among all methods tested, the DNA extraction protocol with glass beads proved to be efficient for P. penetrans and all other species tested (P. crenatus, P. neglectus and P. thornei), generating high quality DNA at comparatively low cost and with a rapid sample throughput.

Item Type: Article
Keywords: diagnostics, DNA yield, glass beads, ITS rRNA, PCR, plant-parasitic nematodes, Pratylenchus crenatus, Pratylenchus neglectus, Pratylenchus penetrans, Pratylenchus thornei, proteinase K
Divisions: Crop and Environment Sciences
Depositing User: Ms Kath Osborn
Date Deposited: 17 Nov 2020 15:13
Last Modified: 17 Nov 2020 15:13
URI: https://hau.repository.guildhe.ac.uk/id/eprint/17593

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