Comparing the efficiency of six common methods for DNA extraction from root-lesion nematodes (Pratylenchus spp.)
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Summary
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.
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Comparing the efficiency of six common methods for DNA extraction from root-lesion nematodes (Pratylenchus spp.)
In: NematologySearch for other papers by Valeria Orlando in
Current site
Google Scholar
PubMed
Search for other papers by Simon G. Edwards in
Current site
Google Scholar
PubMed
Search for other papers by Roy Neilson in
Current site
Google Scholar
PubMed
Search for other papers by Tom Prior in
Current site
Google Scholar
PubMed
Search for other papers by David Roberts in
Current site
Google Scholar
PubMed
Search for other papers by Matthew Back in
Current site
Google Scholar
PubMed
Summary
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.
| All Time | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 1026 | 343 | 40 |
| Full Text Views | 102 | 38 | 5 |
| PDF Views & Downloads | 195 | 72 | 4 |