Pre-mRNA splicing represents an important regulatory layer of eukaryotic gene expression. In the simple budding yeast Saccharomyces cerevisiae, about one-third of all mRNA molecules undergo splicing, and splicing efficiency is tightly regulated, for example, during meiotic differentiation. S. cerevisiae features ...
Read More »A cloud-based workflow to quantify transcript-expression levels in public cancer compendia
Public compendia of sequencing data are now measured in petabytes. Accordingly, it is infeasible for researchers to transfer these data to local computers. Recently, the National Cancer Institute began exploring opportunities to work with molecular data in cloud-computing environments. With ...
Read More »A step-by-step workflow for low-level analysis of single-cell RNA-seq data with Bioconductor
Single-cell RNA sequencing (scRNA-seq) is widely used to profile the transcriptome of individual cells. This provides biological resolution that cannot be matched by bulk RNA sequencing, at the cost of increased technical noise and data complexity. The differences between scRNA-seq ...
Read More »How to standardize the small RNA-Seq workflow for reliable biomarker signatures in the age of liquid biopsies
Small RNA-Seq has emerged as a powerful tool in transcriptomics, gene expression profiling and biomarker discovery. Sequencing cell-free nucleic acids, particularly microRNA (miRNA), from liquid biopsies additionally provides exciting possibilities for molecular diagnostics, and might help establish disease-specific biomarker signatures. ...
Read More »PAR-CLIP and Streamlined Small RNA cDNA Library Preparation Protocol for the Identification of RNA Binding Protein Target Sites
The study of protein-RNA interactions is critical for our understanding of cellular processes and regulatory circuits controlled by RNA binding proteins (RBPs). Recent next generation sequencing-based approaches significantly promoted our understanding of RNA biology and its importance for cell function. ...
Read More »RACE-Seq – extension of human lncRNA transcripts by RACE coupled with long-read high-throughput sequencing
Long non-coding RNAs (lncRNAs) constitute a large, yet mostly uncharacterized fraction of the mammalian transcriptome. Such characterization requires a comprehensive, high-quality annotation of their gene structure and boundaries, which is currently lacking. Here researchers from the Barcelona Institute of Science ...
Read More »Barcodes composed of sample-specific (moderately) degenerate base regions (mDBRs)
Tag-Seq is a high-throughput approach used for discovering SNPs and characterizing gene expression. In comparison to RNA-Seq, Tag-Seq eases data processing and allows detection of rare mRNA species using only one tag per transcript molecule. However, reduced library complexity raises ...
Read More »Recommendations for differential expression analysis and biomarker discovery small RNA-Seq experiments in an age of liquid biopsies
Small RNA-Seq has emerged as a powerful tool in transcriptomics, gene expression profiling and biomarker discovery. Sequencing cell-free nucleic acids, particularly microRNA (miRNA), from liquid biopsies additionally provides exciting possibilities for molecular diagnostics, and might help establish disease-specific biomarker signatures. ...
Read More »Application of an RNA amplification method for reliable single-cell transcriptome analysis
A team led by researchers at the University of Florida present a single-cell RNA amplification approach combining exponential and linear amplification steps using a limited number of PCR cycles and T7-driven in vitro transcription (IVT). Thier approach allows for unlimited ...
Read More »The best RNA-Seq analysis interface…depends on your persective
RNA-sequencing (RNA-seq) has become an established way for measuring gene expression in model organisms and humans. While methods development for refining the corresponding data processing and analysis pipeline is ongoing, protocols for typical steps have been proposed and are widely ...
Read More »