Next-generation Sequencing
Next-generation Sequencing - Related Videos
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Next-generation Sequencing
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2021
The first human genome sequencing project cost $2.7 billion and was declared complete in 2003, after 15 years of international cooperation and collaboration between several research teams and funding agencies. Today, with the advent of next-generation sequencing technologies, the cost and time of sequencing a human genome have dropped over 100 fold. Next-Generation Sequencing Methods Although all next-generation methods use different technologies, they all share a set of standard features.
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Targeted DNA Methylation Analysis by Next-generation Sequencing
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Cited by 83 •
2015
Bisulfite amplicon sequencing (BSAS) is a method for quantifying cytosine methylation in targeted genomic regions of interest. This method uses bisulfite conversion paired with PCR amplification of target regions prior to next-generation sequencing to produce absolute quantitation of DNA methylation at a base-specific level.
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Next-generation Sequencing of 16S Ribosomal RNA Gene Amplicons
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Cited by 119 •
2014
Characterizing microbial community has been a longstanding goal in environmental microbiology. Next-generation sequencing methods now allow for the characterization of microbial communities at an unprecedented depth with minimal cost and labor. We detail here our approach to sequence bacterial 16S ribosomal RNA genes using a benchtop sequencer.
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Collection and Extraction of Saliva DNA for Next Generation Sequencing
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Cited by 45 •
2014
DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.
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Next Generation Sequencing for the Detection of Actionable Mutations in Solid and Liquid Tumors
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Cited by 8 •
2016
This manuscript describes clinical protocols for two next-generation sequencing panels. One panel interrogates hematologic malignancies while the other panel targets genes commonly mutated in solid tumors. Molecular classification of driver mutations in human malignancies offers valuable prognostic and predictive information.
FAQs
Library preparation places fragmented DNA or RNA into a form that can be processed in parallel. The workflow may include amplification or another processing step before sequencing begins. These steps are important because they determine how the original nucleic acid enters the high-throughput readout, allowing the resulting sequences to be aligned and interpreted computationally.
Sequencing cycles translate nucleotide incorporation or other signal-generating reactions into sequence information. Because many molecules are read in parallel, the method can generate large datasets rapidly and support biological analysis at scale. Computational alignment then helps organize the resulting reads for biological interpretation across large datasets.
Compared with traditional sequencing approaches, NGS emphasizes speed, scale, and sensitivity, making large-scale biological analysis more accessible. That distinction matters when the research question extends beyond a small number of sequences, such as examining an entire genome, exome, transcriptome, or defined target set. The chosen scope determines the biological information produced.
After sequencing, computational tools align the resulting sequences and interpret them. This stage connects the raw readout to biological questions, including identifying genetic variation, measuring gene expression, characterizing pathogens, and investigating disease mechanisms. The experiment’s value therefore depends not only on sequence generation but also on how the data are analyzed.
Whole-genome, exome, transcriptome, and targeted sequencing provide different analytical scopes. Researchers can select among them according to whether they need broad genomic information, information from a defined biological component, expression-related data, or focused analysis of selected sequences. This flexibility allows the sequencing design to match the intended biological outcome.
NGS supports genetic variation studies, gene-expression measurement, pathogen characterization, and investigations of disease mechanisms. These applications make it useful across molecular biology and genomics, while its ability to analyze biological material at large scale also contributes to personalized medicine. The resulting data can connect sequence-level observations with broader biological or clinical questions.