The mammalian lung is a highly branched network in which the distal regions of the bronchial tree transform during development into a densely packed honeycomb of alveolar air sacs that mediate gas exchange. Although this transformation has been studied by ...
Read More »Next-Generation Sequencing RNA-Seq Library Construction
This unit presents protocols for construction of next-generation sequencing (NGS) directional RNA sequencing libraries for the Illumina HiSeq and MiSeq from a wide variety of input RNA sources. The protocols are based on the New England Biolabs (NEB) small RNA ...
Read More »Quality assessment and control of tissue specific RNA-seq libraries
RNA-sequencing (RNA-seq) is rapidly emerging as the technology of choice for whole-transcriptome studies. However, RNA-seq is not a bias free technique. It requires large amounts of RNA and library preparation can introduce multiple artifacts, compounded by problems from later stages ...
Read More »6 Changes That’ll Make a Big Difference With Your RNA-seq
from Cofactor Genomics 1. Sample Quality and and stringent QC measures First and foremost, if a sample is of low quality or complexity, both of which can occur due to degradation, then it is best to either not sequence or ...
Read More »AG-NGS: semi-automated preparation of next-generation sequencing libraries using open liquid handling platforms
Next-generation sequencing (NGS) is becoming one of the most widely used technologies in the field of genomics. Library preparation is one of the most critical, hands-on, and time-consuming steps in the NGS workflow. Each library must be prepared in an ...
Read More »Automation of library preparation for RNA -Seq applications
The production of DNA libraries in sufficient quantity and quality is not only the actual sequencing of the essential step in order to achieve meaningful results in accordance with Next Generation Sequencing (NGS ) process . Automated methods must be ...
Read More »Library construction for next-generation sequencing: Overviews and challenges
High-throughput sequencing, also known as next-generation sequencing (NGS), has revolutionized genomic research. In recent years, NGS technology has steadily improved, with costs dropping and the number and range of sequencing applications increasing exponentially. Here, researchers from The Scripps Research Institute ...
Read More »Molecular indexing enables quantitative targeted RNA sequencing and reveals poor efficiencies in standard library preparations
Researchers at Stanford University present a simple molecular indexing method for quantitative targeted RNA sequencing, in which mRNAs of interest are selectively captured from complex cDNA libraries and sequenced to determine their absolute concentrations. cDNA fragments are individually labeled so ...
Read More »Almost all steps of NGS library preparation protocols introduce bias, especially in the case of RNA-Seq
Next-generation sequencing (NGS) has caused a revolution in biology. NGS requires the preparation of libraries in which (fragments of) DNA or RNA molecules are fused with adapters followed by PCR amplification and sequencing. It is evident that robust library preparation ...
Read More »Significant Technical Variations in Low-Input RNA-seq Methodologies
Recent advances in RNA-seq methodologies from limiting amounts of mRNA have facilitated the characterization of rare cell-types in various biological systems. So far, however, technical variations in these methods have not been adequately characterized, vis-à-vis sensitivity, starting with reduced levels ...
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