[一种使用qPCR评估桡足动物染色质衰减值的简单有效方法]。

Genetika Pub Date : 2016-11-01
V I Teterina, Yu A Galimova, N G Sheveleva, L V Sukhanova, T Yu Mayor, A N Zaitseva, S V Kirilchik
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引用次数: 0

摘要

不同种类的淡水类桡足类动物的染色质衰减值(CD)可能存在显著差异。这些差异的生物学和进化作用尚不清楚。为了扩大对这类桡足动物CD分布和大小的认识,需要一种快速评价CD的方法。本研究提出了一种利用实时定量PCR (qPCR)评估桡足类动物CD的简便方法。基因组大小变化的幅度是通过比较目标基因的qPCR片段的荧光曲线来评估的。该方法在四种摆线桡足类动物上进行了试验。独眼羊(Cyclops kolensis)的CD为95.3±1.2;春棘环草为94.6±0.8%;在C. insignis,为82.3±5.2%;首次在巨眼病毒中发现CD,为91.1±2.6%。我们的方法的优点是它的快速,简单和最小的材料要求研究。
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[A simple and effective method for assessing chromatin diminution values in copepods using qPCR].

The value of chromatin diminution (CD) in different species of freshwater cyclopoid copepods can differ significantly. The biological and evolutionary roles of these differences remain unclear. To expand the knowledge on CD distribution and magnitude in this group of copepods, a quick method for its evaluation was required. This study proposes a simple approach for CD assessment in copepods using quantitative realtime PCR (qPCR). The magnitude of changes in the genome size was assessed by comparing fluorescence curves of qPCR fragments of target genes for pre- and post-diminution materials. The method was tested on four cyclopoid copepods species. In Cyclops kolensis, CD was assessed as 95.3 ± 1.2; in Acanthocyclops vernalis it was assessed at 94.6 ± 0.8%; at C. insignis, it was 82.3 ± 5.2%; and for the first time, CD was found in Megacyclops viridis at 91.1 ± 2.6%. The advantages of our approach are its rapidity, simplicity and minimal requirements of materials studied.

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