The attachment of unique molecular identifiers (UMIs) to RNA molecules prior to PCR amplification and sequencing, makes it possible to amplify...
Read More »Quantile normalization of single-cell RNA-seq read counts without unique molecular identifiers
Single-cell RNA-seq (scRNA-seq) profiles gene expression of individual cells. Unique molecular identifiers (UMIs) remove duplicates in read counts resulting from polymerase chain reaction, a major source of noise. For scRNA-seq data lacking UMIs...
Read More »zUMIs: A fast and flexible pipeline to process RNA sequencing data with UMIs
The recent development of sensitive protocols allows to generate RNA-seq libraries of single cells. The throughput of such scRNA-seq protocols is rapidly increasing, enabling the profiling of tens of thousands of cells and opening exciting possibilities to analyse cellular identities. ...
Read More »Gene length and detection bias in single cell RNA sequencing protocols
Single cell RNA sequencing (scRNA-seq) has rapidly gained popularity for profiling transcriptomes of hundreds to thousands of single cells. This technology has led to the discovery of novel cell types and revealed insights into the development of complex tissues. However, ...
Read More »A cost effective 5′ selective single cell transcriptome profiling approach with improved UMI design
Single cell RNA sequencing approaches are instrumental in studies of cell-to-cell variability. 5′ selective transcriptome profiling approaches allow simultaneous definition of the transcription start size and have advantages over 3′ selective approaches which just provide internal sequences close to the ...
Read More »Small-RNA Sequencing of Single Cells
The ability to profile mRNAs in single cells has transformed the molecular identification and the characterization of cell types, states and rare cellular phenotypes. However, current single-cell methods are restricted to long RNAs, primarily mRNAs. Several classes of small RNAs ...
Read More »The impact of amplification on differential expression analyses by RNA-Seq
Currently quantitative RNA-Seq methods are pushed to work with increasingly small starting amounts of RNA that require PCR amplification to generate libraries. However, it is unclear how much noise or bias amplification introduces and how this effects precision and accuracy ...
Read More »