Next Generation Sequencing (NGS) Technologies

Next Generation Sequencing (NGS) Technologies enable a wide variety of methods, allowing researchers to ask virtually any question related to genome, transcriptome, or epigenome of any organism. Sequencing methods differ primarily by how the DNA or RNA samples are obtained and by the data analysis used. The numbers of methods is constantly growing. The most common are small RNA-Seq: Whole Transcriptome Shotgun Sequencing, Exome Sequencing, De Novo Full-Length Transcriptome Analysis and Hybrid Sequencing Approach. Various NGS platforms in the market such as Illumina, SOLiD, and Roche, offer unprecedented ability to apply massively parallel sequencing of transcriptomics (RNA) and genomic (DNA) samples to understand disease and health. The enormous Differential Splicing and RNA Sequencing Data pose a fundamental problem of management and analysis. Various data analysis solutions eliminate the next-generation sequencing data management. ABRF Next-Generation Sequencing (ABRF-NGS) is the most effective technique for accurate sequencing data. For Ribonucleic acid, Single-cell RNA sequencing (scRNA-seq) is carried out which examines the RNA sequence information from individual cells with optimized next generation sequencing (NGS) technologies, providing a higher resolution of cellular differences and a better understanding of the function of an individual cell in the context of its microenvironment.


Whole Transcriptome Shotgun Sequencing is another name for RNA-sequencing and is part of the next-generation sequencing technology. Transcriptome sequencing includes RNA profiling, mRNA transcript expression analysis, and the sequencing and analysis of full-length mRNA transcripts The constantly increasing throughput of next generation sequencing (NGS) technology is revolutionising our ability to describe both genome structure and variation. Recently, numerous draft whole genome sequences have been developed in higher plants, based either in part or entirely on NGS-based sequencing strategies and hybrid sequencing approach is one  of them. De-novo full length Transcriptome analysis recovers more full-length transcripts across a broad range of expression levels, with a sensitivity similar to methods that rely on genome alignments. Exome sequencing can efficiently identify coding variants across a wide range of applications, including population genetics, genetic disease, and cancer studies. RNA sequencing (RNA-Seq) has emerged as a powerful and increasingly cost-effective technology for analysis of transcriptomes. RNA-Seq has several significant advantages over gene expression microarrays, including its high sensitivity and accuracy, broad dynamic range, nucleotide-level resolution, ability to detect novel mRNA transcripts, and ability to analyze pre-mRNA alternative splicing. DNA microarray is used to determine complementary binding of the unknown sequences thus allowing parallel analysis for gene expression and gene discovery. Transcriptome profiling with RNA-Seq is a powerful tool for analyzing gene expression levels of individual samples, as well as for comparing differential gene expression among multiple samples. RNA-Seq also provides information about novel isoforms, including the identification of alternatively spliced transcripts. The goals of the ABRF-NGS is to evaluate the performance of NGS platforms and to identify optimal methods and best practices through the use of reference standards.   The long-term goals of the study include optimizing the detection of genetic variation with the latest sequencing tools; establishing a community resource for self-evaluation and self-improvement that will allow users of NGS technologies to readily compare and improve their own performance data; and facilitating the evaluation of existing and emerging NGS platforms and protocols.

  • RNA-Seq: Whole Transcriptome Shotgun Sequencing
  • Hybrid Sequencing Approach
  • De Novo Full-Length Transcriptome Analysis
  • Exome Sequencing
  • Differential Splicing and RNA Sequencing Data
  • Microarray Gene Expression
  • Transcriptome Databases
  • Transcriptome Profiling by RNA-Seq
  • ABRF Next-Generation Sequencing (ABRF-NGS)
  • Software and Platforms

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