Targeted Ngs
Targeted Ngs - Related Videos
09:18
Mycobacterial DNA Extraction using Bead Beating in Custom Buffer Followed by NGS Workflow
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2025
This protocol shows bead-beating combined with DNA capture bead purification provides a fast and consistent method for extracting DNA from Mycobacterium tuberculosis samples, making it an effective choice for next-generation sequencing applications.
08:38
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.
12:05
RIBO-seq in Bacteria: a Sample Collection and Library Preparation Protocol for NGS Sequencing
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Cited by 5 •
2021
Here we describe the stages of sample collection and preparation for RIBO-seq in bacteria. Sequencing of the libraries prepared according to these guidelines results in sufficient data for comprehensive bioinformatic analysis. The protocol we present is simple, uses standard laboratory equipment and takes seven days from lysis to obtaining libraries.
08:40
Genome-wide Screen for miRNA Targets Using the MISSION Target ID Library
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Cited by 1 •
2012
The Target ID Library is a plasmid-based, genome-wide collection of cloned cDNA used to identify miRNA targets. Here we demonstrate its use and application.
08:56
Recombineering and Gene Targeting
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2023
One of the most widely used tools in modern biology is molecular cloning with restriction enzymes, which create compatible ends between DNA fragments that allow them to be joined together. However, this technique has certain restrictions that limit its applicability for large or complex DNA construct generation. A newer technique that addresses some of these shortcomings is recombineering, which modifies DNA using homologous recombination (HR), the exchange between different DNA molecules based...
FAQs
Probe and primer design determines which genomic regions are enriched before sequencing. Capture-based workflows use designed probes to select target sequences, whereas amplification-based workflows use primers to copy specific regions. Because the assay focuses on predefined loci, design choices directly affect which variants can be observed and how efficiently the selected regions are represented in the sequencing data.
Concentrating sequencing reads on fewer loci increases coverage across the selected regions. Greater coverage can improve analytical sensitivity, meaning the assay is better positioned to detect sequence differences within those targets. This benefit is most relevant when testing a defined set of genes or genomic regions, because sequencing effort is not distributed across the entire genome.
After massively parallel sequencing, bioinformatic alignment compares the generated reads with genomic reference positions. This analysis can identify single-nucleotide changes, insertions, deletions, and copy-number alterations within the selected targets. The resulting variant information is therefore tied to the regions included in the assay rather than to genomic areas outside the predefined design.
Targeted NGS examines a narrower, predefined set of loci, while whole-genome and whole-exome approaches survey substantially broader genomic scopes. The focused design can reduce data-processing demands and direct sequencing depth toward selected regions. In return, findings are limited to the genes or loci included in the target panel, which makes assay scope an important consideration.
A typical workflow begins by defining genomic regions of interest and designing probes or primers for those loci. The target sequences are then captured or amplified, followed by massively parallel sequencing. Bioinformatic alignment subsequently organizes the reads against genomic positions so that sequence variants and copy-number alterations can be assessed within the selected regions.
In genetics, targeted NGS supports disease-gene testing, diagnosis of inherited disorders, and validation of known genomic regions. Cancer profiling is another application, allowing analysis of selected genes or loci relevant to a tumor study. Its focused scope can provide concentrated information while limiting the sequencing and data-processing burden associated with broader genomic assays.