RNA-Seq on libraries made from ERCC external RNA controls, and a mixture of D. melanogaster S2 cell mRNA and ERCC mRNAs. RNA-Seq on libraries made from ERCC external RNA controls, and a mixture of D. melanogaster S2 cell mRNA and ERCC mRNAs

RNA-Seq on libraries made from External RNA Controls Consortium (ERCC) external RNA controls, and a mixture of mRNA from Drosophila melanogaster S2 cell and ERCC mRNAs. We evaluated performance of RNA-Seq on known synthetic PolyA+ mRNAs from the External RNA Controls Consortium (ERCC) alone and in mixtures with PolyA+ mRNA from Drosophila S2 cells. ERCC mRNAs were obtained under Phase V testing from the National Institutes of Standards and Technology (NIST). The ERCC pool contained 96 species of mRNA of various lengths and GC content covering a 2^20 concentration range. Libraries were constructed using 100ng S2 mRNA with 5ng, 2.5ng, or 1ng ERCC mRNAs, and using 50ng ERCC mRNA without S2 cell mRNA. Our data shows an outstanding linear fit between RNA-Seq read density and known input amounts. Overall design: We made libraries with 100ng S2 mRNA with 5ng, 2.5ng or 1ng ERCC mRNAs and with 50ng ERCC mRNAs only. For each library, one lane was sequenced for a 36bp run and around 15 million reads were obtained for each lane.

本数据集针对由外部RNA对照联盟（External RNA Controls Consortium，ERCC）外部RNA对照品，以及黑腹果蝇（Drosophila melanogaster）S2细胞的信使核糖核酸（mRNA）与ERCC mRNA的混合样本构建的文库开展了RNA测序（RNA-Seq）。我们分别针对仅使用ERCC合成的已知聚腺苷酸化mRNA（PolyA+ mRNA），以及其与果蝇S2细胞聚腺苷酸化mRNA的混合体系，评估了RNA测序的检测性能。ERCC mRNA购自美国国家标准与技术研究院（National Institutes of Standards and Technology，NIST）的第五阶段测试批次产品。ERCC对照池涵盖96种不同长度和GC含量的mRNA，其浓度跨度达2^20倍的动态范围。文库构建采用两种方案：一是以100ng果蝇S2 mRNA分别添加5ng、2.5ng或1ng的ERCC mRNA；二是仅使用50ng ERCC mRNA且不添加果蝇S2细胞mRNA。本研究数据显示，RNA测序的读段密度与已知的样本起始投入量之间呈现极佳的线性拟合效果。整体实验设计：我们构建了两类文库：一类为100ng果蝇S2 mRNA分别搭配5ng、2.5ng或1ng ERCC mRNA，另一类为仅含50ng ERCC mRNA的文库。每个文库均通过单泳道进行36bp长度的测序，每条泳道可获得约1500万条读段。