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本研究建立了一种高效、经济的非损伤性扇贝DNA提取方法,在不影响扇贝个体生存状态的前提下,采用擦拭法和室温裂解相结合的方式在20 min内即可获得个体基因组DNA。以虾夷扇贝(Patinopecten yessoensis)、栉孔扇贝(Chlamys farreri)和海湾扇贝(Argopecten irradians)为实验对象,研究了不同擦拭材料(棉签、滤纸)和不同擦拭部位(外套膜、鳃丝、内脏团)的核酸提取效果,评估了本方法在贝类基因分型领域应用的可行性。研究表明,用棉签、滤纸2种材料擦拭扇贝的鳃丝、内脏团或外套膜组织均能获得主带清晰完整的基因组DNA,且不影响扇贝的存活状态。在3种扇贝中采用棉签擦拭法的DNA得率均高于滤纸法,以这2种方式获得的DNA为模板进行SSR和SNP标记分析,能获得均一稳定的扩增条带,SSR标记分型结果准确可靠。本研究建立了简便、快速进行扇贝基因型分析的非损伤性DNA提取方法,为贝类全基因组选育和珍稀贝类资源的种质保护开发提供了新的技术手段。 相似文献
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凡纳滨对虾(Litopenaeusvannamei)是世界也是我国的主导对虾养殖品种,产业的发展离不开良种的支撑,分子育种被认为是加快良种选育的最有效途径,而目标性状相关分子标记的开发是发展分子育种的基础。研究主要目的是建立一种适用于凡纳滨对虾等水产经济物种的高通量候选基因关联分析方法,并在抗弧菌相关标记筛选中进行应用。首次将三代靶向测序技术用于对虾候选基因的基因分型,在抗弧菌性状候选基因LvPI3K的全长序列上发掘到91个SNP位点,通过关联分析鉴定到21个与抗弧菌性状显著相关的SNP标记(P0.05),利用Sanger测序证实了三代靶向测序技术分型结果准确可靠。所建立的基于三代测序的靶向分型方法为凡纳滨对虾等水产动物提供了一种高效、低成本的基因分型方法,所发掘的抗弧菌性状相关位点对开展凡纳滨对虾抗弧菌性状分子育种具有一定指导意义。 相似文献
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《African Journal of Marine Science》2013,35(2):145-153
This study investigated multiple mating in three species of intertidal klipfishes (family Clinidae): super klipfish Clinus superciliosus, bluntnose klipfish Clinus cottoides, and nosestripe klipfish Muraenoclinus dorsalis. These species display the rare reproductive mode of female vivipary with superfetation, a phenomenon where offspring at various stages of development are found within the female. Three or four microsatellite loci were used to genotype 25–30 adults of each species, as well as embryos of 131 C. superciliosus from six broods, 103 C. cottoides from six broods, and 63 M. dorsalis from five broods. The presence of multiple mating by females was assessed by maximum-likelihood sibship reconstruction implemented in the program COLONY, and the non-random distribution of reproductive success among individuals (reproductive skew) within broods was tested using the binomial skew index. Multiple mating by females was pervasive and observed in 83%, 100% and 80% of the broods of C. superciliosus, C. cottoides and M. dorsalis, respectively. The mean number of sires per brood was 4.3, 3.0 and 3.2, and, correspondingly, 5/5, 5/6 and 1/4 of multiply-sired broods showed significant reproductive skew in C. superciliosus, C. cottoides and M. dorsalis, respectively. The presence of pronounced reproductive skew may indicate post-copulatory processes taking place that bias paternity. We compared our results on multiple mating within the klipfish family, and with four other fish families that have vivipary with superfetation, and found the klipfish patterns were among those of live-bearing vertebrate groups with the highest average numbers of sires per brood. 相似文献
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