Normalization of high-throughput small RNA sequencing (sRNA-Seq) data is required to compare sRNA levels across different samples. Commonly used relative normalization approaches can cause erroneous conclusions due to fluctuating small RNA populations between tissues. Researchers from the Austrian Academy of ...
Read More »External calibration with whole-cell spike-ins delivers absolute mRNA fold changes
Gene expression measurements are typically performed on a fixed-weight aliquot of RNA, which assumes that the total number of transcripts per cell stays nearly constant across all conditions. In cases where this assumption does not hold (e.g., when comparing cell ...
Read More »Assessment of single cell RNA-seq normalization methods
UCSD researchers have assessed the performance of seven normalization methods for single cell RNA-seq using data generated from dilution of RNA samples. Their analyses showed that methods considering spike-in ERCC RNA molecules significantly outperformed those not considering ERCCs. This work ...
Read More »Using Synthetic Mouse Spike-In Transcripts to Evaluate RNA-Seq Analysis Tools
One of the key applications of next-generation sequencing (NGS) technologies is RNA-Seq for transcriptome genome-wide analysis. Although multiple studies have evaluated and benchmarked RNA-Seq tools dedicated to gene level analysis, few studies have assessed their effectiveness on the transcript-isoform level. ...
Read More »Assessing technical performance in differential gene expression experiments with external spike-in RNA control ratio mixtures
There is a critical need for standard approaches to assess, report and compare the technical performance of genome-scale differential gene expression experiments. Here, an international team led by researchers at the NIST assess technical performance with a proposed standard ‘dashboard’ ...
Read More »RNA-Seq Blog – Poll Results
We asked: Do you use spike in controls in your RNA-Seq experiments? 65 Respondents Thanks to all who participated! Check out the latest poll question on the RNA-Seq Blog homepage.
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