The preparation of small RNA cDNA sequencing libraries depends on the unbiased ligation of adapters to the RNA ends. Small RNA with 5' recessed ends are poor substrates for enzymatic adapter ligation, but this 5' adapter ligation problem can...
Read More »A novel method for the capture-based purification of whole viral native RNA genomes
Current technologies for targeted characterization and manipulation of viral RNA primarily involve amplification or ultracentrifugation with isopycnic gradients of viral particles to decrease host RNA background. The former strategy is non-compatible for characterizing properties innate to RNA strands such as ...
Read More »STARmap – three-dimensional intact-tissue sequencing of single-cell transcriptional states
Retrieving high-content gene-expression information while retaining three-dimensional (3D) positional anatomy at cellular resolution has been difficult, limiting integrative understanding of structure and function in complex biological tissues. Stanford...
Read More »Optimized methodology for the generation of RNA-sequencing libraries from low-input starting material
RNA sequencing (RNA-seq) has become an important tool for examining the role of the transcriptome to biological processes. While RNA-seq has been widely adopted as a popular approach in many experimental designs, from gene discovery to mechanistic validation of targets, ...
Read More »Systematic comparison of small RNA library preparation protocols for next-generation sequencing
Next-generation sequencing technologies have revolutionized the study of small RNAs (sRNAs) on a genome-wide scale. However, classical sRNA library preparation methods introduce serious bias, mainly during adapter ligation steps. Several types...
Read More »An efficient RNAseq library prep method for determining reverse transcription termination sites
Researchers from the Scripps Research Institute describe a method for making Illumina-compatible sequencing libraries from RNA. This protocol can be used for standard RNAseq analysis for detecting differentially expressed genes. In addition, this protocol is ideally suited for adapting to ...
Read More »Incorporation of unique molecular identifiers (UMIs) in TruSeq adapters improves the accuracy of quantitative sequencing
Quantitative analysis of next-generation sequencing (NGS) data requires discriminating duplicate reads generated by PCR from identical molecules that are of unique origin. Typically, PCR duplicates are identified as sequence reads that align to the same genomic coordinates using reference-based alignment. ...
Read More »Poly-A selection or Ribo-depletion?
RNA sequencing (RNA-seq) has become an indispensable tool to identify disease associated transcriptional profiles and determine the molecular underpinnings of diseases. However, the broad adaptation of the methodology into the clinic is still hampered by inconsistent results from different RNA-seq ...
Read More »Preparation of highly multiplexed small RNA sequencing libraries
MicroRNAs (miRNAs) are ~22-nucleotide-long small non-coding RNAs that regulate the expression of protein-coding genes by base pairing to partially complementary target sites, preferentially located in the 3´ untranslated region (UTR) of target mRNAs. The expression and function of miRNAs have ...
Read More »SomaGenics, Inc. launched RealSeq®-AC, a novel kit for preparing small-RNA sequencing libraries
July 2017, – SomaGenics, Inc. launched RealSeq®-AC, a novel kit for preparing small-RNA sequencing libraries that eliminates incorporation bias in Next-Generation-Sequencing (NSG). Previously commercially available small RNA sequencing kits’ incorporation biases leads to over/under estimating the abundance of some miRNAs, ...
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