Delghandi Madjid, Mortensen Atle, Westgaard Jon-Ivar
Department of Aquaculture, Norwegian Institute of Fisheries and Aquaculture Research (Fiskeriforskning), Muninbakken 9-13, Breivika, NO-9291 Tromsø, Norway.
Mar Biotechnol (NY). 2003 Mar-Apr;5(2):141-8. doi: 10.1007/s10126-002-0086-x.
A novel hexaplex assay system including Gmo8, Gmo19, Gmo35, Gmo37, Tch11, and Tch12 microsatellites from Atlantic cod, consisting of trinucleotide or tetranucleotide repeat units, is introduced. All 6 loci were coamplified in a single reaction employing dye-labeled primers. Alleles from these loci were sized using an internal standard by automated sample processing in an ABI 310 Genetic Analyser. Amplified alleles in profiles containing selected microsatellites were typed clearly, providing easily interpretable results. Sequencing data indicated that alleles at all loci consisted of simple repeat units. This may help minimize the likelihood of stuttering upon polymerase chain reaction amplification. The results suggest that the presented hexaplex assay system may be a useful tool in a selective breeding program in which genetic identification will allow different genotypes to be reared together from fertilization. This should have a great impact as it will make selective breeding more efficient.
介绍了一种新型的六重检测系统，该系统包含来自大西洋鳕鱼的Gmo8、Gmo19、Gmo35、Gmo37、Tch11和Tch12微卫星，由三核苷酸或四核苷酸重复单元组成。所有6个位点在一个反应中使用染料标记引物进行共扩增。通过在ABI 310遗传分析仪中进行自动样品处理，使用内标对这些位点的等位基因进行大小测定。在包含选定微卫星的图谱中，扩增的等位基因分型清晰，提供易于解释的结果。测序数据表明，所有位点的等位基因均由简单重复单元组成。这可能有助于将聚合酶链反应扩增时出现口吃的可能性降至最低。结果表明，所提出的六重检测系统可能是选择性育种计划中的一个有用工具，在该计划中，基因鉴定将使不同基因型从受精开始就一起饲养。这应该会产生很大影响，因为它将使选择性育种更有效。
