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Primer GX2

引物GX2

Company: Illumina
Catalog#: 1000592
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Preparation of cDNA Library for dRNA-seq
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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 preparation of degradome RNA complementary DNA library for high-through-put sequencing (dRNA-seq) using Illumina GA II sequencing platform, which is currently most popular and cost-effective. Using this ... [摘要]  微小RNA(miRNA)是真核生物体中基因表达的普遍存在的调节剂,其指导Argonaute蛋白(AGO)基于序列互补性切割靶mRNA或抑制其翻译。在植物中,miRNA定向切割发生在靶mRNA上,在约5至11个核苷酸(nt)上游至miRNA的5'端结合的位点。 miRNA定向切割产物(降解组)的测序广泛用作验证miRNA介导的调节的生物信息学预测和鉴定已知miRNA的新靶标的方法。在这里,我们描述了使用Illumina GA II测序平台制备高通量测序(dRNA-seq)的降解组RNA互补DNA文库的方案,目前这是最受欢迎和成本效益的。使用这个协议,我们成功地使用三个茄科植物,包括烟草,番茄和马铃薯生成三个dRNA seq图书馆。尽管该协议是用单链适配器开发的,但应该能够通过用多路复用兼容的3'衔接子代替3'衔接子并用索引引物代替PCR引物来产生多重文库。

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