UDI ADAPTERS FOR MULTIPLEXING ILLUMINA® SEQUENCING LIBRARIES

NEXTFLEX® Unique Dual Index Barcodes

384 UDI adapters for mitigating sample misassignment due to index hopping

  • Unique dual index adapters for multiplexing up to 384 Illumina® sequencing libraries
  • Mitigate index hopping or spread of signal that can occur during sequencing
  • Each lot of the NEXTFLEX® Unique Dual Index barcodes is functionally validated and tested for index purity by sequencing
  • Compatible with both paired-end and single-read Illumina® sequencing

For labs who want greater multiplexing capacity, PerkinElmer also offers a set of 1,536 NEXTFLEX UDI barcodes.

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For research use only. Not for use in diagnostic procedures.

  • NOVA-514150

       $2,241

    BARCODES 1-96
    192 rxns

  • NOVA-514151

       $2,241

    BARCODES 97-192
    192 rxns

  • NOVA-514152

       $2,241

    BARCODES 193 – 288
    192 rxns

  • NOVA-514153

       $2,241

    BARCODES 289 – 384
    192 rxns

Data Security in Sequencing Applications

The NEXTFLEX® Unique Dual Index Barcodes are barcoded adapters, for sequencing on Illumina® platforms, that provide unprecedented data security in sequencing applications. Multiplexing with NEXTFLEX® Unique Dual Index barcodes drastically increases scale while reducing costs by allowing the user to pool multiple libraries in a single flow cell lane. These barcoded adapters can be used with single read or paired-end read sequencing, and are compatible with NEXTFLEX® library prep kits designed for Illumina® sequencing, or other workflows for Illumina® sequencing that involve the ligation of adapters to adenylated fragments.

UDIs Prevent Data Ambiguity Caused by Index Hopping

The NEXTFLEX® Unique Dual Index Barcodes utilize two unique 8 nt indexes on each UDI adapter. The 8 nt indices are designed with a Hamming Distance of at least three throughout the entire series, permitting error correction. Each NEXTFLEX® Unique Dual Index Barcode comprises the entire flow cell binding site, sequencing primer binding site, and index sequences, making PCR enrichment optional. Patterned flow cells, such as those found on the NovaSeq® and HiSeq® 3000/4000 platforms have been recognized to suffer from increased rates of sample mis-assignment during sequencing. The use of adapters with unique dual indices during sequencing prevents such mis-assigned reads from appearing in final data sets, allowing for the highest assurance of data integrity.

Reduce Run-to-Run Sample Carryover & Sample Crosstalk with UDIs

Even users who don’t routinely perform high-throughput multiplexing benefit from the superior design of the NEXTFLEX® Unique Dual Index Barcodes. The ability to avoid using the same set of indices consecutively reduces concerns about run-to-run sample carryover and sample crosstalk. They are designed to specifically address the index hopping, or spread of signal, phenomenon associated with Illumina® platforms that utilize a patterned flow cell.

KIT CONTENTS

  • NEXTFLEX® Unique Dual Index Barcodes* (25 µM)
  • NEXTFLEX® Primer Mix (12.5 µM)

* The UDI barcodes are oriented column-wise on each plate (1-8, 9-16, etc)
For automation single-use plates, NEXTFLEX® Unique Dual Index Barcodes may be at a different specified concentration. Please inquire for additional details.

Selected Citations that Reference the Use of the NEXTFLEX UDI Barcodes:

  • Estermann, M. (2020). Mouse embryonic stem cells self-organize into trunk-like structures with neural tube and somites. doi:10.1242/prelights.18906.
  • Gardner, E. J., Prigmore, E., Gallone, G., Danecek, P., Samocha, K. E., Handsaker, J., . . . Hurles, M. E. (2019). Contribution of retrotransposition to developmental disorders. Nature Communications, 10(1). doi:10.1038/s41467-019-12520-y.
  • Gaeta, N. C., Bean, E., Miles, A. M., Daniel Ubriaco Oliveira Gonçalves De Carvalho, Alemán, M. A., Carvalho, J. S., . . . Ganda, E. (2020). A Cross-Sectional Study of Dairy Cattle Metagenomes Reveals Increased Antimicrobial Resistance in Animals Farmed in a Heavy Metal Contaminated Environment. Frontiers in Microbiology, 11. doi:10.3389/fmicb.2020.590325.
  • Hirose, K., Chang, S., Yu, H., Wang, J., Barca, E., Chen, X., . . . Huang, G. N. (2019). Loss of a novel striated muscle-enriched mitochondrial protein Coq10a enhances postnatal cardiac hypertrophic growth. doi:10.1101/755793.
  • Leon, K. E., et al. (2020) DOT1L modulates the senescence-associated secretory phenotype through epigenetic regulation of IL1A. bioRxiv 2020.08.21.258020; doi: 10.1101/2020.08.21.258020.
  • Miura, H., Takahashi, S., Shibata, T. et al. Mapping replication timing domains genome wide in single mammalian cells with single-cell DNA replication sequencing. Nat Protoc 15, 4058–4100 (2020). https://doi.org/10.1038/s41596-020-0378-5
  • Starr, T. N., Greaney, A. J., Hilton, S. K., Crawford, K. H., Navarro, M. J., Bowen, J. E., . . . Bloom, J. D. (2020). Deep mutational scanning of SARS-CoV-2 receptor binding domain reveals constraints on folding and ACE2 binding. doi:10.1101/2020.06.17.157982.
  • Veenvliet, J. V., et al. (2020) Mouse embryonic stem cells self-organize into trunk-like structures with neural tube and somites. bioRxiv 2020.03.04.974949; doi: 10.1101/2020.03.04.974949
Q: Are the NEXTFLEX® unique dual index barcodes sequence-verified for purity?

A: Yes, each lot of the adapters undergoes stringent QC procedures that include verification of the index sequence purity by sequencing on a Illumina® MiSeq® instrument.


Q: Are the NEXTFLEX® unique dual index barcodes functionally validated?

A: Yes, each lot of these adapters undergoes stringent QC procedures that include functional validation of the barcoded adapter by making a library.


Q: What is the lead time when I order the NEXTFLEX® unique dual index barcodes?

A: They are in-stock and ready-to-ship.


Q: Why are the NEXTFLEX® unique dual index barcodes only supplied at two reactions of each barcode in the 96-well plate?

A: The NEXTFLEX® unique dual index barcodes are supplied in aliquots of two reactions of each barcode to reduce the potential for contamination, which increases with repeated use of the same plate.


Q: Must I perform paired-end sequencing with NEXTFLEX® Unique Dual Index Barcodes?

A: No, they are compatible with paired-end and single-read sequencing runs. The I7 and I5 indexes are read in separate sequencing events than Read 1 and Read 2.


Q: Can I use the NEXTFLEX® Unique Dual Index Barcodes in single index experiments?

A: Yes, you can configure your sequencing run to only read the I7 index when you set up the run.

Each NEXTFLEX® Unique Dual Index Barcodes Kit contains 96 unique dual-index barcodes, enabling users to multiplex up to 384 samples per flow cell lane. Each lot of the NEXTFLEX® Unique Dual Index Barcodes is functionally validated and tested for index purity by sequencing. NEXTFLEX® Unique Dual Index Barcodes are supplied in 96-well plates with each well containing 2 reactions of each barcode. These kits ship on dry ice. The shelf life of each reagent is at least two years when stored properly.

For automation single-use plates, NEXTFLEX® Unique Dual Index Barcodes will be at a different specified concentration and each well will contain 1 reaction of each barcode.  Please inquire for additional details. These kits ship on dry ice.

IMPORTANT: We have recently modified the sequence of 13 indexes of the 1-384 Unique Dual Barcode sets.

If you have acquired your barcodes before December 11th 2020, or you just want to access to the previous version please contatct us: [email protected]

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For research use only. Not for use in diagnostic procedures.