Beaufort Genetics Research Programme, School of Biological, Earth and Environmental Sciences/Aquauculture and Fisheries Development Centre, University College Cork, Distillery Fields, North Mall, Cork, Ireland.
J Fish Biol. 2013 Mar;82(3):944-58. doi: 10.1111/jfb.12034. Epub 2013 Jan 30.
By combining next-generation sequencing technology (454) and reduced representation library (RRL) construction, the rapid and economical isolation of over 25 000 potential single-nucleotide polymorphisms (SNP) and >6000 putative microsatellite loci from c. 2% of the genome of the non-model teleost, Atlantic cod Gadus morhua from the Celtic Sea, south of Ireland, was demonstrated. A small-scale validation of markers indicated that 80% (11 of 14) of SNP loci and 40% (6 of 15) of the microsatellite loci could be amplified and showed variability. The results clearly show that small-scale next-generation sequencing of RRL genomes is an economical and rapid approach for simultaneous SNP and microsatellite discovery that is applicable to any species. The low cost and relatively small investment in time allows for positive exploitation of ascertainment bias to design markers applicable to specific populations and study questions.
通过结合下一代测序技术(454)和简化代表性文库(RRL)构建,成功快速且经济地从爱尔兰南部凯尔特海的非模式硬骨鱼大西洋鳕鱼(Gadus morhua)的基因组中约 2%的区域中分离出超过 25000 个潜在的单核苷酸多态性(SNP)和 >6000 个假定的微卫星位点。对标记的小规模验证表明,80%(14 个中的 11 个)的 SNP 位点和 40%(15 个中的 6 个)的微卫星位点可以扩增并表现出可变性。结果清楚地表明,RRL 基因组的小规模下一代测序是一种经济快速的 SNP 和微卫星发现方法,适用于任何物种。这种方法的低成本和相对较少的时间投入,允许积极利用确定偏差来设计适用于特定群体和研究问题的标记。