Low-cost and High-throughput RNA-seq Library Preparation for Illumina Sequencing from Plant Tissue
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Author:
Date:
2020-10-20
[Abstract] Transcriptome analysis can provide clues to biological processes affected in different genetic backgrounds or/and under various conditions. The price of RNA sequencing (RNA-seq) has decreased enough so that medium- to large-scale transcriptome analyses in a range of conditions are feasible. However, the price and variety of options for library preparation of RNA-seq can still be daunting to those who would like to use RNA-seq for their first time or for a single experiment. Among the criteria for selecting a library preparation protocol are the method of RNA isolation, nucleotide ...
[摘要] [摘要] 转录组分析可以为不同遗传背景或不同条件下的生物学过程提供线索。RNA测序(RNA-seq)的价格已经下降到足够低的程度,因此在各种条件下进行中大规模转录组分析是可行的。然而,对于那些希望第一次使用RNA-seq或进行单个实验的人来说,RNA-seq库制备的价格和各种选择仍然是令人望而生畏的。选择文库制备方案的标准包括RNA分离方法、核苷酸片段化以获得所需的大小范围,以及文库索引以汇集测序样本进行多路复用。在这里,我们提出了一个高质量和高通量的选择,从多聚腺苷酸mRNA制备文库用于转录组分析。高质量和高通量的方案选择都包括通过磁珠使poly-A尾部沉淀,cDNA合成,然后通过Tn5介导的“标记”同时裂解和添加适配器的步骤。该方案的所有步骤均已通过拟南芥叶片和幼苗组织的验证,并简化为协同工作,在资金和时间上成本最低,因此旨在为转录组分析提供一个初学者友好的从开始到完成的RNA序列库制备。 [背景] 通过Southern印迹、expressed sequence ...
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Preparation of cDNA Library for dRNA-seq
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Author:
Date:
2012-12-05
[Abstract] microRNAs (miRNAs) are ubiquitous regulators of gene expression in eukaryotic organisms, which guide Argonaute proteins (AGO) to cleave target mRNA or inhibit its translation based on sequence complementarity. In plants, miRNA directed cleavage occurs on the target mRNA at about 10 to 11 nucleotide (nt) up stream to the site where the 5’ end of miRNA binds. Sequencing of the miRNA directed cleavage products (degradome) is widely employed as a way to both verify bioinformatic predictions of miRNA mediated regulation and identify novel targets of known miRNAs. Here we describe a protocol for ...
[摘要] 该实验方案的中文版正在准备中...
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