Project description:Array hybridizations profiling diverse human tissues; for each array the human tissue RNA is hybridized against a universal reference RNA. Experiments 1-3 are replicates of universal reference RNA vs. genomic DNA hybridizations, used to estimate transcript abundance. Arrays 4-8 are tissue RNA vs. genomic DNA hybridizations, carried out to evaluate the method for estimating transcript abundance (see manuscript for details). Logical set Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: other
Project description:nCounter miRNA Expression Assay data for all profiled miRQC samples prepared by the miRQC Consortium. In this study, we systematically compared 12 commercially available miRNA expression platforms by measuring an identical set of standardized positive and negative control samples, including human universal reference RNA, human brain RNA and titrations thereof, human serum samples, and synthetic spikes from miRNA family members with varying homology. We developed novel and robust quality metrics to objectively assess platform performance of very different technologies such as small RNA sequencing, RT-qPCR and (microarray) hybridization. We assessed reproducibility, sensitivity, accuracy, specificity, and concordance of differential expression.
Project description:Array hybridizations profiling diverse human tissues; for each array the human tissue RNA is hybridized against a universal reference RNA. Series_type: Logical set Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: other
Project description:We have developed two methods for efficiently consructing RNA-seq libraries using transposition. Each method constructs high quality RNA-seq libraries when compared to standard approaches. One of the methods (Directional Tn-RNA-seq) maintains strand-of-origin information and exhibits strand specificity comparable to current approaches. RNA-seq libraries were constructed from ECC-1, a human cell line, and Universal Human Reference RNA using transposon-based and standard RNA-seq library construciton methods.
Project description:A large number of computational methods have been recently developed for analyzing differential gene expression (DE) in RNA-seq data. We report on a comprehensive evaluation of the commonly used DE methods using the SEQC benchmark data set and data from ENCODE project. We evaluated a number of key features including: normalization, accuracy of DE detection and DE analysis when one condition has no detectable expression. We found significant differences among the methods. Furthermore, computational methods designed for DE detection from expression array data perform comparably to methods customized for RNA-seq. Most importantly, our results demonstrate that increasing the number of replicate samples significantly improves detection power over increased sequencing depth. The Sequencing Quality Control Consortium generated two datasets from two reference RNA samples in order to evaluate transcriptome profiling by next-generation sequencing technology. Each sample contains one of the reference RNA source and a set of synthetic RNAs from the External RNA Control Consortium (ERCC) at known concentrations. Group A contains 5 replicates of the Strategene Universal Human Reference RNA (UHRR), which is composed of total RNA from 10 human cell lines, with 2% by volume of ERCC mix 1. Group B includes 5 replicate samples of the Ambion Human Brain Reference RNA (HBRR) with 2% by volume of ERCC mix 2. The ERCC spike-in control is a mixture of 92 synthetic polyadenylated oligonucleotides of 250-2000 nucleotides long that are meant to resemble human transcripts.
Project description:Fibroblast cells were cultured from human skin, either tumor (basal cell carcinoma) or non-tumor tissue. Cells were propagated for several passages in culture, then switched to low (0.1%) serum conditions for 48 hours prior to mRNA harvest. Reference for all arrays was Stratagene Universal Human Reference RNA. Set of arrays organized by shared biological context, such as organism, tumors types, processes, etc. Keywords: Logical Set
Project description:Both spotted long oligonucleotide arrays and Affymetrix GeneChips were used to measure differential gene expression in two RNA samples (K562 erythroleukemia RNA and the Stratagene Universal Human Reference RNA). For Affymetrix technology, the two RNAs were analyzed separately. There are two replicates for K562 RNA (GSM4843 and GSM4844) and three for the Stratagene Universal reference (GSM4845-GSM4847). Two types of spotted long oligonucleotide arrays were used. These arrays were used to do two-color hybridizations to directly compare K562 and Universal reference RNAs. One array (GPL273) was made with probes from the Operon Human Genome Oligo Set Version 1. These arrays were used with unamplified cDNA probes (6 replicates, GSM4848-GSM4853), with aRNA probes produced by one round of T7 RNA polymerase-based amplification (6 replicates, GSM4854-GSM4859), and with aRNA probes produced by two rounds of amplification (2 replicates, GSM4860-GSM4861). Keywords: parallel sample
Project description:We identified human small RNAs containing m1A (N1-methyladenosine) by m1A RIP and TGIRT-seq. Small RNA-seq and long RNA-seq was performed after TRMT6/61A knock-down.
Project description:nCounter miRNA Expression Assay data for all profiled miRQC samples prepared by the miRQC Consortium. In this study, we systematically compared 12 commercially available miRNA expression platforms by measuring an identical set of standardized positive and negative control samples, including human universal reference RNA, human brain RNA and titrations thereof, human serum samples, and synthetic spikes from miRNA family members with varying homology. We developed novel and robust quality metrics to objectively assess platform performance of very different technologies such as small RNA sequencing, RT-qPCR and (microarray) hybridization. We assessed reproducibility, sensitivity, accuracy, specificity, and concordance of differential expression. miRQC Consortium multi-platform comparision. *This represents the NanoString component only