NEXTflex-96™ DNA Barcodes

Illumina-compatible adapters for multiplexing DNA-seq libraries

Catalog# Product Name Quantity US List Price
NOVA-514105 NEXTflex-96™ DNA Barcodes (in 96-well plate) 768 rxns $7,133 Buy Now
NOVA-514106 NEXTflex-96™ DNA Barcodes (in tubes) 768 rxns $7,133 Buy Now


  • Compatible with NEXTflex® Rapid DNA-Seq, NEXTflex® Methyl DNA-Seq Kit and other DNA-Seq applications
  • 8 nt index contained within adapter sequence
  • Considerably reduce your Illumina per-sample sequencing cost by multiplexing DNA-Seq samples
  • Increase your sequencing scale by pooling several samples on a single flow cell
  • Supplied in 96-well plate format or 96 microfuge tubes
  • Compatible with Illumina® next-generation sequencing platforms


The NEXTflex-96™ DNA Barcodes are compatible with Illumina® sequencing and provide flexibility and high-throughput capabilities in sequencing applications. They significantly increase scale while reducing costs by allowing the user to pool multiple library preparations in a single flow cell lane. These adapters compatible with Illumina® sequencing utilize an indexed adapter with an 8 nt unique sequence. This allows for proper differentiation between samples, preventing poor reads caused by single base errors introduced during PCR. The NEXTflex index is contained within the adapter sequence, eliminating the need to perform PCR to add flow cell binding sequences. The NEXTflex-96 DNA Barcode index sequences are designed to have a hamming distance of 4, allowing discrimination of different barcodes even in the case of an error in the barcode read.

These adapters are supplied in either a 96-well format (cat # 514105) or in microfuge tubes (cat # 514106). These barcodes can be used with single, paired-end, and multiplex reads and are compatible with the NEXTflex Rapid DNA-Seq Kit, the NEXTflex Methyl DNA-Seq Kitthe NEXTflex Cell Free DNA-Seq and other genomic DNA library prep protocols. 

Do you want to index your samples but don’t want to use 96 different barcodes? We also offer the NEXTflex DNA Barcodes in sets of 6, 12, 24, and 48 barcodes. 

Do you want to multiplex more than 96 samples? The new 

NEXTflex-HT™ Barcodes are ideal for researchers who want to multiplex up to 384 samples with single-index adapters.

NEXTflex RNA-Seq Barcodes and NEXTflex-96 RNA-Seq Barcodes are also available for your RNA-Seq multiplexing needs.

A PDF containing the indexes contained within the NEXTflex-96 DNA Barcodes can be downloaded here. Additionally, Excel files containing the index sequences are also available upon request.

Avoiding registration failure with low level multiplexing

Registration failure could occur if the color balance was not maintained between the red and green lasers (used to sequence A/C bases and G/T bases, respectively). Read our blog post, Tech Tips – Barcode Recommendations for Low Level Multiplexing, to learn how to avoid registration failure on an Illumina® sequencer caused by lack sufficient index sequence diversity. 

Select Publications that Cite Use of the NEXTflex-96 DNA Barcodes: 

Amaroa, F., et al. (2016) Genetic characterization of Arrabida virus, a novel phlebovirus isolated in South Portugal. Virus Research. doi:10.1016/j.virusres.2016.01.004.

David, L. A., et al. (2015) Gut Microbial Succession Follows Acute Secretory Diarrhea in Humans. 6:3, doi: 10.1128/mBio.00381-15. 

Derboven, E., Ekker, H., Kusenda, B., Bulankova, P., Riha K. (2014) Role of STN1 and DNA Polymerase α in Telomere Stability and Genome-Wide Replication in Arabidopsis. PLoS Genetics. DOI: 10.1371/journal.pgen.1004682.

Devall, M., et al. (2015) A comparison of mitochondrial DNA isolation methods in frozen post-mortem human brain tissue—applications for studies of mitochondrial genetics in brain disorders. Biotechniques 59(4): 241-246.

Henry, I. et al. (2014) Efficient Genome-Wide Detection and Cataloging of EMS-Induced Mutations Using Exome Capture and Next-Generation Sequencing. Plant Cell.

Ivansson, E. L., et al. (2016) Variants within the SP110 nuclear body protein modify risk of canine degenerative myelopathy. PNAS PLUS. doi:10.1073/pnas.1600084113.

Jiang, L. et al. (2014) ZBED6 Modulates the Transcription of Myogenic Genes in Mouse Myoblast Cells. PLoS ONE 9(4): e94187. doi: 10.1371/journal.pone.0094187.

Kis, O. et al. (2017) Circulating tumour DNA sequence analysis as an alternative to multiple myeloma bone marrow aspirates. Nat. Commun. 8, 15086 doi: 10.1038/ncomms15086.

Kloth, M., et al. (2015) Activating ERBB2/HER2 mutations indicate susceptibility to pan-HER inhibitors in Lynch and Lynch-like colorectal cancer. Gut doi:10.1136/gutjnl-2014-309026.

Maheshwari, S. et al. (2015) Naturally Occurring Differences in CENH3 Affect Chromosome Segregation in Zygotic Mitosis of Hybrids. PLoS Genet. 11(1) doi:  10.1371/journal.pgen.1004970

Ravi, M., et. al.  (2014) A haploid genetics toolbox for Arabidopsis thaliana. Nature Communications. 5, 5334 doi: 10.1038/ncomms6334.

Thakur J. and Sanyal K. (Feb 2013) Efficient neocentromere formation is suppressed by gene conversion to maintain centromere function at native physical chromosomal loci in Candida albicans. Genome Research. doi: 10.1101/gr.141614.112

Yamane A. et al. (2013) RPA Accumulation during Class Switch Recombination Represents 5′–3′ DNA-End Resection during the S–G2/M Phase of the Cell Cycle. Cell Reports.

Zastrow-Hayesa, G. M., et al. (2015) Southern-by-Sequencing: A Robust Screening Approach for Molecular Characterization of Genetically Modified Crops. The Plant Genome. doi:10.3835/plantgenome2014.08.0037.

Kit Specs

The NEXTflex-96 DNA Barcodes Kits contain 8 reactions each of 96 unique barcodes, enabling the user to multiplex up to 96 samples. This kit is shipped on dry ice.

Kit Contents

NEXTflex-96 HT Barcode Adapter (25 µM)
NEXTflex Primer Mix (12.5 µM)

NEXTflex-96™ Adapter Design

DNASeq Adapter Illumina Compatible

The NEXTflex-96™ adapters contain the full complement of sequencing flow cell binding regions (A, B), which eliminates the need to perform PCR to add the barcode tag.