Experimental Design for Time-Series RNA-Seq Analysis of Gene Expression and Alternative Splicing.

Experimental Design for Time-Series RNA-Seq Analysis of Gene Expression and Alternative Splicing.
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基因表达和选择性剪接的时间序列 RNA-Seq 分析的实验设计。

DOI:
10.1007/978-1-0716-1912-4_14
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发表时间:
2022
期刊:
Methods in molecular biology (Clifton, N.J.)
影响因子:
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通讯作者:
Tzioutziou NA
Tzioutziou NA
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Tzioutziou NA

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RNA测序(RNA-seq)是目前分析差异基因表达的首选方法。为了充分利用全基因组转录方法产生的丰富数据,实验的初始设计至关重要。自然界中的生物节律无处不在,由内源性基因网络驱动,统称为生物钟。测量昼夜节律基因的表达需要时程实验,这些实验考虑了影响表达水平变异性的一天中的时间因素。我们在这里描述了一种方法,用于描述植物在经历降温过程中的表达和选择性剪接的日变化。该方法使用廉价的日常实验室设备,并利用RNA-SEQ应用程序(3D RNA-SEQ),该应用程序可以处理复杂的实验设计,并且几乎不需要先前的生物信息学专业知识。
RNA-sequencing (RNA-seq) is currently the method of choice for analysis of differential gene expression. To fully exploit the wealth of data generated from genome-wide transcriptomic approaches, the initial design of the experiment is of paramount importance. Biological rhythms in nature are pervasive and are driven by endogenous gene networks collectively known as circadian clocks. Measuring circadian gene expression requires time-course experiments which take into account time-of-day factors influencing variability in expression levels. We describe here an approach for characterizing diurnal changes in expression and alternative splicing for plants undergoing cooling. The method uses inexpensive everyday laboratory equipment and utilizes an RNA-seq application (3D RNA-seq) that can handle complex experimental designs and requires little or no prior bioinformatics expertise.
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发表时间: 2000-12-15
期刊: SCIENCE
影响因子: 56.9
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