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基于 Illumina 测序技术的斜带石斑鱼基因组调查和多态性微卫星标记的开发。

Genome survey and development of polymorphic microsatellite loci for Sillago sihama based on Illumina sequencing technology.

机构信息

Guangdong Provincial Key Laboratory of Marine Biotechnology, Institute of Marine Sciences, Shantou University, 243 Daxue Road, Shantou, 515063, China.

STU‑UMT Joint Shellfish Research Laboratory, Shantou University, Shantou, 515063, China.

出版信息

Mol Biol Rep. 2020 Apr;47(4):3011-3017. doi: 10.1007/s11033-020-05348-z. Epub 2020 Mar 2.

Abstract

In this study, we first conducted a genome survey assay for Sillago sihama by Illumina sequencing platform, and then developed 15 polymorphic microsatellite loci in a wild population. A total of 129.46 Gb raw data were obtained, of which 115.07 Gb were clean data, with a sequencing depth of 179.3-folds. This genome was estimated to be 522.6 Mb in size, with the heterozygosity, repeat content and GC content being 0.63%, 21% and 44%. A total of 630,028 microsatellites were identified from the genome, of which, dinucleotide repeat was the most abundant (56.80%), followed by mononucleotide repeat (30.23%). Furthermore, 60 pairs of primers were designed and synthesized based on microsatellite sequences, of which 15 were polymorphic in a wild population. A total of 91 alleles were found, with an average of 6.07 per locus. Number of alleles, observed and expected heterozygosity per locus ranged from two to 13, from 0.250 to 0.862, and from 0.396 to 0.901, respectively. Twelve loci were highly informative (PIC > 0.5), and the others were medium informative (0.25 < PIC < 0.5). Seven loci deviated from Hardy-Weinberg equilibrium after Bonferroni correction (P < 0.0033). No significant linkage disequilibrium was detected between loci pairs. This study provided a large number of genomic resources and 15 polymorphic microsatellite loci that should be helpful for the further genetic studies in S. sihama.

摘要

在本研究中,我们首先使用 Illumina 测序平台对灰鲷进行了基因组调查分析,然后在野生种群中开发了 15 个多态性微卫星标记。共获得了 129.46GB 的原始数据,其中 115.07GB 为清洁数据,测序深度为 179.3 倍。该基因组大小估计为 522.6Mb,杂合度、重复含量和 GC 含量分别为 0.63%、21%和 44%。从基因组中鉴定出 630028 个微卫星,其中二核苷酸重复最为丰富(56.80%),其次是单核苷酸重复(30.23%)。此外,基于微卫星序列设计并合成了 60 对引物,其中 15 对在野生种群中具有多态性。共发现 91 个等位基因,每个位点平均 6.07 个。每个位点的等位基因数、观测和预期杂合度范围为 2 至 13,0.250 至 0.862,0.396 至 0.901。12 个位点高度信息丰富(PIC>0.5),其余为中度信息丰富(0.25<PIC<0.5)。经过 Bonferroni 校正后,有 7 个位点偏离 Hardy-Weinberg 平衡(P<0.0033)。未检测到位点对之间存在显著的连锁不平衡。本研究提供了大量的基因组资源和 15 个多态性微卫星标记,这对进一步研究灰鲷的遗传结构应该是有帮助的。

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