• Single-index adapters for multiplexing up to 384 Illumina libraries
  • Increase your sequencing scale by pooling hundreds of samples on a single flow cell
  • Compatible with both paired-end and single-read Illumina® sequencing
  • Maximum flexibility of suitable pooling combinations for Illumina® platforms
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For research use only. Not for use in diagnostic procedures.

  • NOVA-514174


    BARCODES 1-96
    (in 96-well plate)

  • NOVA-514175


    BARCODES 97-192
    (in 96-well plate)

  • NOVA-514176


    BARCODES 193-288
    (in 96-well plate)

  • NOVA-514177


    BARCODES 289-384
    (in 96-well plate)



    BARCODES 385-2,380
    (in 96-well plate)

Ideal for High-Throughput Capabilities for Sequencing Applications

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

Dual Error Correction Prevents Ambiguity

The NEXTFLEX-HT Barcodes utilize a single-index adapter containing a 12 nt unique sequence. The 12 nt indices are designed with a Hamming Distance of at least five throughout the entire series of 2380, permitting dual error correction. Dual error correction enables proper differentiation between samples, by preventing ambiguity from PCR errors or sequencing instrument miscalling. The NEXTFLEX-HT index is contained within the adapter sequence, making PCR enrichment optional.

Reduce Run-to-Run Sample Carryover & Sample Crosstalk

Even users who don’t routinely perform high-throughput multiplexing benefit from the superior design of the NEXTFLEX-HT Barcodes. The ability to avoid using the same set of indices consecutively reduces concerns about run-to-run sample carryover and sample crosstalk. Plus, each pair of consecutive barcodes are fully color balanced and suitable for two-plex sequencing.


  • NEXTFLEX-HT Barcode Adapter (25 µM)
  • NEXTFLEX® Primer Mix (12.5 µM)


  • Sequences of NEXTFLEX-HT Barcode Indexes – Excel / PDF

The NEXTFLEX-HT Barcodes Kits contain 6 or 96 unique single-index barcodes, enabling users to multiplex up to 384 samples per flow cell lane. NEXTFLEX-HT Barcodes – 6 are supplied in tubes containing 8 reactions worth of each barcode. The sets of 96 NEXTFLEX-HT Barcodes are supplied in 96-well plates with each well containing 2 reactions of each barcode. These kits ship on dry ice.

What is the concentration of the NEXTFLEX-HT Barcodes?
The concentration of the NEXTFLEX-HT Barcodes is 25 µM.

Are these barcodes compatible with both paired-end and single-read Illumina® sequencing?
Yes, the NEXTFLEX-HT Barcodes can be used with both SR and PE sequencing.

Do the NEXTFLEX-HT Barcodes Kits contain PCR primers?
Yes, these barcodes are supplied with PCR primers.

At what concentration are the NEXTFLEX® PCR primers supplied?
The NEXTFLEX® PCR Primers concentration is 12.5 µM.

Can I order more NEXTFLEX® PCR primers alone?
Yes, the NEXTFLEX® PCR Primers are available in sets of 48, 96, 192, 384 or 768 reactions. Contact [email protected] for more information.

How is the quality of the NEXTFLEX-HT Barcodes assured?
Each lot of the NEXTFLEX® barcodes is sequence verified on an Illumina® MiSeq® instrument.

What type of purification is used when the NEXTFLEX® DNA Barcodes are synthesized?
In order to ensure the highest quality sequencing data, the NEXTFLEX-HT Barcodes are HPLC-purified, minimizing the risk of mispriming events due to truncations and other data quality-compromising errors.

Are the NEXTFLEX-HT Barcodes methylated?
No they are not methylated; therefore they are not compatible with bisulfite-seq applications. However, they are compatible with all other DNA-Seq applications including MeDIP and MeCAP sequencing. The NEXTFLEX® Bisulfite-Seq Barcodes are provided for library prep requiring methylated adapters.

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