Next Generation Sequencing
Support and Educational Content

NGS detection of low frequency genetic variants using novel, molecular sequencing adapters

Precision medicine for oncology requires accurate and sensitive molecular characterization. However, sample degradation, and polymerase and sequencing errors reduce accuracy for sequencing genetic variants.

This webinar is an excellent resource for both new and experienced investigators interested in analyzing low-frequency variants using NGS methods. In this presentation, Dr Mirna Jarosz discusses the challenges encountered when looking for variants against a complex genetic background. She presents a method developed by IDT scientists for producing the necessary high read depth, while demonstrating a 300-fold reduction in false positives at or above 0.25% variant frequency. The discussion includes a ranked overview of error correction approaches, and why unique molecular adapters are especially important for low-frequency variant analysis.

In this presentation, you will also learn about the basic workflow for probe-based target capture using xGen® Lockdown® Probes. These sequences are individually synthesized and quality checked, and outperform other commercially available options by providing deep, uniform coverage of targeted regions. Dr Jarosz describes the use of xGen Blocking Oligos to minimize hybridization artifacts and, thereby, improve on-target performance.


Product focus: Target capture reagents

xGen® Lockdown® Probes—target capture probe pools for NGS

xGen Lockdown Probes are pools of individually synthesized, quality controlled, and normalized hybridization probes. Use them to generate custom capture panels for targeted sequencing to enhance the performance of existing panels. xGen Predesigned Gene Capture Probe Pools are available for any human RefSeq coding gene. Select from predesigned and custom probes that offer:

  • Sensitive detection of SNPs, indels, CNV, LOH, and translocations
  • GMP compliance for clinical and diagnostics research
  • Flexibility to augment existing panels or create completely custom panels
  • Quick delivery

Discover more about xGen Lockdown Probes.


xGen Lockdown Panels

xGen Lockdown Panels are preconfigured, validated, and stocked pools of xGen Lockdown Probes for targeted next generation sequencing of defined gene families:

  • xGen Exome Research Panel
  • xGen Acute Myeloid Leukemia Panel
  • xGen Pan-Cancer Panel
  • xGen Inherited Diseases Panel
  • xGen Human ID Research Panel
  • xGen Human mtDNA Research Panel

Discover more about xGen Lockdown Panels.


xGen Lockdown Reagents—hybridization and wash kit

xGen Lockdown Reagents have been optimized to deliver deep, even coverage of targets captured using xGen Lockdown Probes and Panels. Achieve uniform coverage with hybridization and wash buffers that are optimized for target enrichment using xGen Lockdown Probes and Panels. A short, 4-hour hybridization protocol generates results quickly.

Discover more about xGen Lockdown Reagents.


xGen Blocking Oligos

xGen Blocking Oligos prevent adapter cross-hybridization and minimize off-target capture, increasing specificity during targeted sequencing. Universal Blocking Oligos—TS Mix are designed for use with Illumina barcoded adapters. This ready-to-use mix effectively blocks dual- or single-index adapters to significantly improve on-target performance.

Also available are:

  • xGen Standard Blocking Oligos
  • xGen Custom Blocking Oligos
  • xGen Blocking Oligos for diagnostics—contact GMPinfo@idtdna.com for information
  • xGen Blocking Oligos for Ion Torrent adapters

Discover more about xGen blocking Oligos.

Additonal reading

How important are those NGS metrics?—Of the many metrics used in evaluating target capture data for NGS applications, read which ones our researchers consider important for evaluating performance of target enrichment panels.

Towards providing personalized medicine—considerations for reliable data—Read how scientists at Geneseeq Technology, Inc. improved their target capture methods to increase accuracy in clinical diagnostics by using optimized blocking oligos and stringent hybridization conditions.

Advantages of high quality, probe-based gene capture panels—Target enrichment by hybrid capture lets you focus your genomic analysis on specific regions of interest, increasing depth of coverage of targeted sequences and improving the detection of rare genomic events. Create your own custom human gene capture panels quickly and cost-effectively using IDT preconfigured pools of probes targeting the coding sequences (CDS) of human protein-coding genes, or with predesigned disease or exome panels.


Review other DECODED Online newsletter articles on NGS applications.


You can also browse our DECODED Online newsletter for additional application reviews, lab tips, and citation summaries to facilitate your research.


Author: Hans Packer, PhD, is a scientific writer at IDT.

© 2017 Integrated DNA Technologies. All rights reserved. Trademarks contained herein are the property of Integrated DNA Technologies, Inc. or their respective owners. For specific trademark and licensing information, see www.idtdna.com/trademarks.


xGen® Blocking Oligos

Adapter blocking oligos increase the number of on-target reads by preventing non-specific binding during hybridization.

Find blockers for your platform ≫


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Improving Uniform Coverage of Targeted Sequences for NGS

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