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利用 MiSeq 技术开发和鉴定丰年虾(鳃足纲:无甲目)微卫星引物。

Development and characterization of microsatellite primers for Triops granarius (Branchiopoda: Notostraca) using MiSeq technology.

机构信息

Department of Medical Education Education City, Qatar Foundation, Weill Cornell Medicine - Qatar, 24144, Doha, Qatar.

Natural History Museum, University of Oslo, Sars Gate 1, 0562, Oslo, Norway.

出版信息

Mol Biol Rep. 2022 Oct;49(10):10121-10125. doi: 10.1007/s11033-022-07804-4. Epub 2022 Sep 4.

DOI:10.1007/s11033-022-07804-4
PMID:36057875
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9515016/
Abstract

BACKGROUND

Next-generation sequencing technology has allowed for the rapid development of microsatellites, neutral polymorphic markers that can be used for the analysis of population structure.

METHODS AND RESULTS

In this study, we performed whole-genome sequencing using the Illumina MiSeq system and de novo assembly to design microsatellite primers for Triops granarius populations in Qatar. The developed microsatellites are suitable for future studies of genetic structuring among geographically isolated freshwater pools. A total of 23 different primer pairs produced typical microsatellite results, with each pair successfully amplified in up to 40 individuals. Only five of the loci produced a significant departure from Hardy-Weinberg equilibrium.

CONCLUSIONS

Some of the underlying mechanisms regarding the few loci that deviated from HWE may be further investigated to determine the source of deviation. As T. granarius is the most widely distributed species of the family, the development of these molecular markers would be useful for conducting population genetics and biogeographical studies broadly.

摘要

背景

新一代测序技术使得微卫星的快速发展成为可能,微卫星是中性多态标记,可以用于分析种群结构。

方法和结果

在这项研究中,我们使用 Illumina MiSeq 系统进行全基因组测序和从头组装,为卡塔尔的丰年虾种群设计微卫星引物。开发的微卫星适用于未来对地理隔离的淡水池塘之间遗传结构的研究。总共 23 对不同的引物产生了典型的微卫星结果,每对引物最多可成功扩增 40 个个体。只有 5 个位点偏离 Hardy-Weinberg 平衡有显著差异。

结论

对于偏离 HWE 的少数几个位点,可能需要进一步研究一些潜在机制,以确定偏离的来源。由于丰年虾是该科分布最广的物种,因此开发这些分子标记将有助于广泛开展种群遗传学和生物地理学研究。

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QDD version 3.1: a user-friendly computer program for microsatellite selection and primer design revisited: experimental validation of variables determining genotyping success rate.QDD版本3.1:一个用于微卫星选择和引物设计的用户友好型计算机程序再探讨:决定基因分型成功率的变量的实验验证
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An economic method for the fluorescent labeling of PCR fragments.一种用于PCR片段荧光标记的经济方法。
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