NEXTFLEX® PCR-Free Barcodes

  • 6 nt index contained within adapter sequence eliminates the need to perform PCR to add flow cell binding sequences
  • Up to 48 multiplexed samples (up to 384 reactions per kit)
  • Compatible with NEXTFLEX® PCR-Free DNA Sequencing Kit
  • Considerably reduce your per-sample sequencing cost by barcoded multiplexing
  • Increase your sequencing scale by pooling 100s of samples on a single flow cell
  • Compatible with Illumina® Next-Generation Sequencing platforms
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For research use only. Not for use in diagnostic procedures.

  • NOVA-514110


    6 BARCODES | 48 RXNS

  • NOVA-514111


    12 BARCODES | 96 RXNS

  • NOVA-514112


    24 BARCODES | 192 RXNS

  • NOVA-514113


    48 BARCODES | 384 RXNS

Pool Multiple Library Preparations in a Single Flow Cell Lane

NEXTFLEX® PCR-Free Barcodes are barcoded adapters for multiplexing Illumina® libraries which 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. The NEXTFLEX® PCR-Free Barcodes accomplish this by using an indexed adapter with a 6 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. These barcodes can be used with single, paired-end, and multiplex reads and are compatible with the NEXTFLEX® PCR-Free Sequencing Kits and other PCR-Free DNA library prep protocols.


  • NEXTFLEX® PCR-Free Barcode Adapter (50 µM)


  • Sequences of NEXTFLEX® PCR-Free Barcode Indexes – Excel / PDF

Selected Publications that Reference Using the NEXTFLEX® PCR-Free DNA Barcodes: 

  • Bartoli, C., Carrere, S., Lamichhane, J. R., Varvaro, L. and Cindy E. Morris, C.  E. (2015) Genome Sequencing of 10 Pseudomonas syringae Strains Representing Different Host Range Spectra. Genome Announc. 3: e00379-15.
  • Chusova, O, et al. (2014) Effect of pine bark on the biotransformation of trinitrotoluene and on the bacterial community structure in a batch experiment. Ecological Technology. Vol. 35, Issue 19.
  • Chusova, O. et al. (2015) Biotransformation of pink water TNT on the surface of a low-cost adsorbent pine bar. Biodegradation. 1 – 12.
  • Evrony, Cai, et al. (2012) Single-Neuron Sequencing Analysis of L1 Retrotransposition and Somatic Mutation in the Human Brain. Cell. Vol. 151, Issue 3, pp. 483-496.
  • Hasbún, R., Iturra, C., Bravo, S., Rebolledo-Jaramillo, B. and Valledor, L. (2016) Differential Methylation of Genomic Regions Associated with Heteroblasty Detected by M&M Algorithm in the Nonmodel Species Eucalyptus globulus Labill. International Journal of Genomics. 4395153. doi: 10.1155/2016/4395153.
  • Kawahara-Miki R., Sano S., Nunome M. et al. (2013) Next-generation sequencing reveals genomic features in the Japanese quail. Genomics. DOI: 10.1016/j.ygeno.2013.03.006.
  • Kocher, A., et al. (2016) Vector soup: high-throughput identification of neotropical phlebotomine sand flies using metabarcoding. Molecular Ecology Resources. doi:10.1111/1755-0998.12556.
  • Kofler, R., Nolte, V. and Schlötterer, C. (2015) Tempo and Mode of Transposable Element Activity in Drosophila. PLOS Genetics. doi: 10.1371/journal.pgen.1005406.
  • Kofler, R., Nolte, V. and Schlötterer, C. (2015) The impact of library preparation protocols on the consistency of allele frequency estimates in Pool-Seq data. Molecular Ecology Resources. doi: 10.1111/1755-0998.12432.
  • Ligi, T, et al. (2013) Characterization of bacterial communities in soil and sediment of a created riverine wetland complex using high-throughput 16S rRNA amplicon sequencing. Ecological Engineering. DOI: 10.1016/j.ecoleng.2013.09.007.
  • Mändar, R. et al. (2015) Complementary seminovaginal microbiome in couples. Research in Microbiology. doi:10.1016/j.resmic.2015.03.009.
  • Mathys J., Vos C. et al. (2013) RNAseq-based transcriptome analysis of Lactuca sativa infected by the fungal necrotroph Botrytis cinerea. Plant, Cell & Environment. DOI: 10.1111/pce.12106.
  • Mazueta, C., Bouchierb, C. and Popoffadoi, M. R. (2015) Draft Genome Sequence of Clostridium botulinum Strain 277-00 Type B2. Genome Announc. 3:2 e00211-15. doi: 10.1128/genomeA.00211-15.
  • Petersen, G., et al. (2015) Phylogeny of the Alismatales (Monocotyledons) and the relationship of Acorus (Acorales?). Cladistics. doi: 10.1111/cla.12120.
  • Sato S, Sesay AK, Holder AA (2013) The Unique Structure of the Apicoplast Genome of the Rodent Malaria Parasite Plasmodium chabaudi chabaudi. PLoS ONE 8(4): e61778. doi:10.1371/journal.pone.0061778.
  • Smidt, I., et al. (2015) Comparison of detection methods for vaginal lactobacilli. Beneficial Microbes. In press.
  • Suzuki, S, et al. (2014) Physiological and genomic features of highly alkaliphilic hydrogen-utilizing Betaproteobacteria from a continental serpentinizing site. Nature Communications 5, (3900).
  • Tiirik, K, et al., (2014) Characterization of the bacterioplankton community and its antibiotic resistance genes in the Baltic Sea. Biotechnology and Applied Biochemistry. Vol. 61, Issue 1, pp. 23–33.
  • Won, HH, et al. (2013) Detecting somatic genetic alterations in tumor specimens by exon capture and massively parallel sequencing. J. Vis. Exp. (80), e50710, doi:10.3791/50710.

What is the concentration of the NEXTFLEX® PCR-Free DNA Barcodes?
The NEXTFLEX® PCR-Free DNA Barcodes concentration is 50 µM.

Do the NEXTFLEX® PCR-Free DNA Barcodes Kits contain PCR primers?
No, these barcodes are not supplied with PCR primers are they are designed to be used with the NEXTFLEX® PCR-Free DNA-Seq Kit.

How is the quality of the NEXTFLEX® PCR-Free DNA Barcodes ensured?
Each lot of the NEXTFLEX® PCR-Free DNA Barcodes are sequence verified on an Illumina® MiSeq® instrument.

What type of purification is used when the NEXTFLEX® PCR-Free DNA Barcodes are synthesized?
The NEXTFLEX® Barcodes are HPLC-purified to ensure high quality sequencing data is obtained. HPLC-purified barcodes minimize the risk of mispriming events due to truncations and other errors that may compromise sequencing data data quality.

The NEXTFLEX® PCR-Free Barcodes Kits contains 6, 12, 24 or 48 unique barcodes, enabling the user to multiplex up to 48 samples per flow cell lane. These kits ship on dry ice.

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