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QIAseq Index Kits

Indices for QIAseq Targeted NGS now available in unique dual bar-coded format
  • Optimized design exclusively from QIAGEN
  • QC of manufacturing lots by NGS
  • Tube and plate formats available for convenience and scalable sample throughput
  • Unique Dual Index adapters available for multiplexing up to 192 samples with QIAseq Targeted NGS solutions*

* Not suitable for QIAseq library prep kits that utilize Y-shaped adapter styles. See our QIAseq FX DNA Library Kits for information on QIAseq Unique Dual Index Y-Adapter Kits.

 

QIAseq Targeted Panels enable Sample to Insight next-generation sequencing (NGS) of DNA or RNA. These optimized solutions facilitate ultrasensitive variant and clonotype detection from cells, tissue and biofluids using integrated Unique Molecular Indices (UMIs).

In order to de-multiplex pooled libraries and assign reads to the appropriate original source library, NGS workflows require the addition of an adapter complex that includes both a common nucleotide sequence, as well as sample index oligonucleotides.

Compatible only with Illumina instruments, our Unique Dual Indexes (UDIs) are optimized for QIAseq Targeted Panel workflows, including QIAseq Targeted DNA, QIAseq RNAscan and QIAseq Immune Repertoire RNA Library Kits. Exclusively available from QIAGEN for QIAseq targeted NGS applications, QIAseq UDI Kits provide the highest level of process safety and ensure confidence in sequencing data by mitigating “index hopping” and mis-assignment of reads. The combination of both UMIs and UDIs provide QIAseq Targeted Panels with a remarkable level of accuracy. Combinatorial, non-unique dual index adapters are also available.

Need a quote for your research project or would you like to discuss your project with our specialist team? Contact Us

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Cat No./ID: 333714
QIAseq 12-Index I (48)
963,00 €
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Box containing combinatorial dual-indexed adapters, for indexing up to 12 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 48 samples

Cat No./ID: 333715
QIAseq 8-Unique Dual Index Set A (48)
1 208,00 €
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Box containing unique dual-indexed adapters, for indexing up to 8 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 48 samples; 1st of 2 sets required for multiplexing up to 16 samples

Cat No./ID: 333716
QIAseq 8-Unique Dual Index Set B (48)
1 208,00 €
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Box containing unique dual-indexed adapters, for indexing up to 8 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 48 samples; 2nd of 2 sets required for multiplexing up to 16 samples

Cat No./ID: 333725
QIAseq 96-Unique Dual Index Set A (384)
5 206,00 €
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Box containing unique dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 1st of 2 sets required for multiplexing up to 192 samples

Cat No./ID: 333727
QIAseq 96-Index I Set A (384)
4 245,00 €
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Box containing combinatorial dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 1st of 4 sets required for multiplexing 384 samples

Cat No./ID: 333735
QIAseq 96-Unique Dual Index Set B (384)
5 206,00 €
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Box containing unique dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 2nd of 2 sets required for multiplexing up to 192 samples

Cat No./ID: 333737
QIAseq 96-Index I Set B (384)
4 245,00 €
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Box containing combinatorial dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 2nd of 4 sets required for multiplexing 384 samples

Cat No./ID: 333747
QIAseq 96-Index I Set C (384)
4 245,00 €
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Box containing combinatorial dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 3rd of 4 sets required for multiplexing 384 samples

Cat No./ID: 333757
QIAseq 96-Index I Set D (384)
4 245,00 €
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Box containing combinatorial dual-indexed adapters, for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Illumina platforms; enough to process a total of 384 samples; 4th of 4 sets required for multiplexing 384 samples

Cat No./ID: 333764
QIAseq 12-Index L (48)
963,00 €
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Box containing oligos for indexing up to 12 samples for QIAseq Targeted Panel sequencing on Ion Torrent platforms; enough to process a total of 48 samples

Cat No./ID: 333777
QIAseq 96-Index L (384)
4 245,00 €
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Box containing oligos for indexing up to 96 samples for QIAseq Targeted Panel sequencing on Ion Torrent platforms; enough to process a total of 384 samples

QIAseq Index Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
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Product Details

9
Principle of molecular barcodes

A variant identified in a sample represents one of two events: a true or false variant. False variants can be introduced at any step during the workflow, including sequencing reactions. This results in the inability to accurately and confidently call rare variants (those present at 1% of the sample). Due to PCR duplicates generated in amplification steps, all DNA fragments look exactly the same, and there is no way to tell whether a specific DNA fragment is a unique DNA molecule or a duplicate of a DNA molecule. With molecular barcodes, since each unique DNA molecule is barcoded before any amplification takes place, unique DNA molecules are identified by their unique barcodes, and PCR duplicates carrying the same barcode are removed, thereby increasing the sensitivity of the panel.

9
Uniformity

The QIAseq targeted DNA panels deliver outstanding sequencing metrics. 6000 SNPs were enriched from 20 ng of NA12878 DNA. Library was constructed for sequencing on a MiSeq, with 4,000,000 reads generated. The panel achieved a uniformity of 99.5% at 0.2x of mean coverage, and 96% at 0.5x of mean coverage.

9
Coverage of GC-rich genomic regions

The QIAseq DNA panels use a proprietary buffer mixture to efficiently sequence GC-rich regions within the genome. Two examples are shown here: CEBPA and CCND1. Complete coverage of exonic regions within those two genes is achieved.

9
Workflow

Isolated DNA, as low as 20 ng, is enzymatically fragmented to generate small pieces of dsDNA. This is followed by the library construction step, in which IL-N7 adapters, molecular barcodes, and sample indexes are incorporated into DNA fragments generated in the previous step. Library fragments now serve as templates for target enrichment using single primer extension. In this step, targets are enriched using a single gene-specific primer and a universal forward primer. The final step is library amplification and sample indexing (for dual indexing) using the IL-S5 sample index primer and a universal primer.

Performance
  • Accuracy: Innovative digital sequencing (incorporating molecular barcodes) eliminates PCR duplication and amplification artifacts to detect low-frequency variants with high confidence (see figure Principle of molecular barcodes).
  • Specificity: The unique combination of our proprietary primer design algorithm and rigorous testing of every primer assay guarantees high specificity and accurate results.
  • Uniformity: The QIAseq Targeted DNA Panel workflow has been optimized to deliver highly uniform sequencing results, to ensure sequencing capacity is utilized very efficiently (see figure Uniformity).
  • Sensitivity: Digital DNA sequencing approach is optimized to deliver high confidence in calling low-frequency DNA variants. Over 90% sensitivity for 1% NA12878 SNP and indel on typical coding region with false positive less than 15 per mega base region when variants are detected with tiled primer design to cover complete coding region of each gene.
  • Universality: The chemistry used in the QIAseq Targeted DNA Panels and workflow is compatible with both regular and GC-rich genomic regions, allowing one to achieve 100% coverage of genes rich in GC content such as CEBPA and CCND1 (see figure Coverage of GC-rich genomic regions).
  • Flexibility: The QIAseq Targeted DNA Panels offer a high degree of flexibility in content and sample multiplexing. Several cataloged panels have been developed for a wide range of applications. One can also build a custom panel for a specific content, or extend the contents of an existing cataloged panel. Up to 384 samples can be multiplexed using the QIAseq indices.
Principle

PCR duplicates are a major issue in targeted DNA sequencing, since, through PCR amplification, they turn unique DNA molecules into identical DNA molecules that cannot be distinguished from each other. In addition, errors from PCR amplification and sequencing process may also be present in final reads that lead to false-positive variants in sequencing results. This, in turn, results in the inability to confidently call DNA variants present at low frequencies in the starting DNA material. To overcome the issue of PCR duplicates and amplification artifacts, the QIAseq Targeted DNA Panels use digital sequencing by incorporating molecular barcodes into the starting DNA material before any amplification takes place, thereby preserving the uniqueness of the starting DNA molecules and overcoming the issues of PCR duplicates, false positives and library bias.

Procedure

The entire workflow of the QIAseq Targeted DNA Panels to go from extracted DNA to sequencing-ready libraries can be completed in 9 hours (see figure Workflow). Extracted DNA is fragmented, genomic targets are molecularly barcoded and enriched, and libraries are constructed. Sequencing files can be fed into the QIAseq pipeline, a cloud-based data analysis pipeline, which will filter, map and align reads, as well as count unique molecular barcodes associated with targeted genomic regions, and call variants with a barcode-aware algorithm. This data can then be fed into IVA or QCI for interpretation.

Applications

QIAseq Targeted DNA Indices can be used for indexing samples enriched with either QIAseq Targeted DNA Panels or QIAseq Targeted RNAscan Panels, and are available for both Illumina and Ion Torrent platforms.

Product Resources

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