Increasing the Efficiency of Microsatellite Discovery from Poorly Enriched Libraries in Coniferous Forest Species |
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Authors: | Scott LJ Cross M Shepherd M Maguire T Henry RJ |
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Institution: | (1) Cooperative Research Centre for Sustainable Production Forestry, Centre for Plant Conservation Genetics, Southern Cross University, Lismore, NSW, 2480, Australia;(2) Centre for Coastal Management and Centre for Plant Conservation Genetics, Southern Cross University, Australia |
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Abstract: | Microsatellites are PCR based markers widely applicable in many plant species. Although the discovery and development of new microsatellites is a costly and technically demanding process, they are widely used in many plant systems. In coniferous tree species, the large genome size and repetitive DNA regions further aggravate the process, often resulting in libraries with very low enrichment efficiencies. Such species have proven intractable to efficient microsatellite discovery. For example, enriched microsatellite libraries produced for seven angiosperm species had enrichment efficiencies up to 50%, whereas libraries for three conifers, generated concurrently, under identical conditions, all had enrichment efficiencies below 10%. This report describes a simple strategy for recovering useful microsatellites from two of these conifer libraries, Pinus elliottii and Araucaria cuninghamii. Enrichment efficiencies increased from 0% to 100% and 1.5% to 98%, respectively. The strategy utilised DIG labelled oligo probes to identify clones containing microsatellite inserts. The technique is cost effective and rapid, utilising basic techniques and equipment that are widely available. |
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Keywords: | Araucaria enrichment forestry Pinus SSR tree species |
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