NEXTflex® Rapid DNA-Seq Library Prep Kit for Illumina® Sequencing

DNA-seq library prep for Illumina® sequencing platforms

Catalog# Product Name Quantity US List Price
NEXTflex® Rapid DNA Sequencing Kit
8 rxns
Buy Now
NOVA-5144-02 NEXTflex® Rapid DNA Sequencing Kit 48 rxns $1,238 Buy Now
NOVA-5144-03 NEXTflex® Rapid DNA-Seq Kit Bundle with DNA Barcodes 1 - 24
48 rxns $961 Buy Now
NOVA-5144-04 NEXTflex® Rapid DNA-Seq Kit Bundle with DNA Barcodes 25 - 48
48 rxns $961 Buy Now
  Barcodes for Use with Less Than 10 ng of DNA

NEXTflex® ChIP-seq Barcodes - 6 48 rxns $240 Buy Now
NEXTflex® ChIP-seq Barcodes - 12
96 rxns $474 Buy Now
NEXTflex® ChIP-seq Barcodes - 24
192 rxns $937 Buy Now
NEXTflex® ChIP-seq Barcodes - 48
384 rxns $1,845 Buy Now
NEXTflex-96™ ChIP-seq Barcodes
768 rxns $3,567 Buy Now
   Barcodes for Use with 10 ng or more of DNA      
NOVA-514101 NEXTflex® DNA Barcodes - 6 48 rxns $480 Buy Now
NOVA-514102 NEXTflex® DNA Barcodes - 12 96 rxns $947 Buy Now
NOVA-514103 NEXTflex® DNA Barcodes - 24 192 rxns $1,876 Buy Now
NOVA-514104 NEXTflex® DNA Barcodes - 48 384 rxns $3,690 Buy Now
NOVA-514105 NEXTflex® DNA Barcodes - 96 768 rxns $7,133 Buy Now
NEXTflex Rapid Illumina DNA-Seq Library Prep Kit
  • Flexible amounts of input DNA from 1 ng to 1 µg
  • Fast workflow requiring only 2 hours, with minimal hands-on time
  • Enhanced Adapter Ligation Technology offers a larger number of unique sequencing reads
  • Automation-friendly workflow
  • Flexible barcode options – up to 384 single-index barcodes available
  • Compatible with Illumina® sequencing platforms including the MiSeq®, HiSeq®, NextSeq® and HiSeq® X Ten
  • Prepare single, paired-end and multiplexed genomic DNA libraries
  • Automation protocols are available for the PerkinElmer Sciclone NGS and NGSx Workstation and the  Biomek® FX and Biomek® FXP Laboratory Automation Workstations

Bioo Scientific’s NEXTflex™ Rapid DNA-Seq Kit is a rapid DNA library prep kit producing libraries for Illumina® sequencing from as little as 1 ng of DNA in two hours. This kit is ideal for use with genomic DNA, FFPE samples, ChIP DNA, upstream target capture samples, pooled amplicons and low input clinical samples. This DNA-Seq library prep kit is highly flexible in terms of sample requirements, accommodating a range of input DNA amounts from 1 ng to 1 µg. The optimized protocol allows for fast generation of non-size-selected libraries and size-selected libraries using reproducible gel-free double SPRI selection. 

The NEXTflex Rapid DNA-Seq Kit incorporates Enhanced Adapter Ligation Technology, which facilitates ligation of long adapters, resulting in longer and more diverse sequencing reads. Bioo Scientific’s NEXTflex Ligation and Polymerase reaction mixes ensure the highest quality libraries for superior performance. Study results in the white paper, Optimizing Library Preparation: Enhanced Adapter Ligation Technology, demonstrate the superior ligation efficiency of the NEXTflex Rapid DNA-Seq Library Prep Kit for creating NGS libraries for Illumina® sequencing.


Rapid DNA Library Prep Workflow

Cost Effective NEXTflex Rapid DNA-Seq Kit Bundles

To simplify your DNA sequencing Bioo Scientific now offers affordable bundles containing NEXTflex Rapid DNA-Seq library prep reagents and NEXTflex DNA Barcodes. There are two different versions of this kit. They each contain the 48 rxn NEXTflex Rapid DNA-Seq Kit and 24 unique NEXTflex DNA Barcodes (in aliquots of 2 reactions each).

Flexible Multiplexing Options

The NEXTflex Adapters are long, annealed adapters containing indexed sequences that offer an improved multiplexing workflow and flexible setup. These barcodes can be used with single, paired-end and multiplex reads. Bioo Scientific strives to make available flexible barcoding options. The NEXTflex Rapid DNA-Seq Kit is designed to be used with the NEXTflex™ DNA BarcodesNEXTflex-96™ DNA Barcodes and NEXTflex-HT™ Barcodes when 10 ng or more of starting material is used. These barcodes are available in sets of 6, 12, 24, 48, 96 and 384 unique indexed adapters.

When less than 10 ng of starting material is available, the NEXTflex™ ChIP-Seq Barcodes and NEXTflex-96™ ChIP-Seq Barcodes should be used. These barcodes are available in sets of 6, 12, 24, 48, and 96 unique indexed adapters. 

Automation Compatibility

The NEXTflex Rapid DNA-Seq Library Prep Kit was designed for easy migration onto automated liquid handling platforms. Currently methods are available for the Beckman-Coulter Biomek® FX and Biomek® FXP Laboratory Automation Workstations. Additionally a validated automation protocol for use with the PerkinElmer Sciclone G3 NGS platform is now available. Download the Sciclone NGS and NGSx Workstation Automation Guides for the NEXTflex Rapid DNA-Seq Kit.

Publications that Cite Using the NEXTflex Rapid DNA-Seq Kit


Brodie, J., et al. (2016) Characterising the microbiome of Corallina officinalis, a dominant calcified intertidal red alga. FEMS Microbiology Ecology. doi: 10.1093/femsec/fiw110.

Campana, M. G., et al. (2014) False positives complicate ancient pathogen identifications using high-throughput shotgun sequencing. BMC Research Notes. 7:111


Campbella, M. A., et al. (2015) Genome expansion via lineage splitting and genome reduction in the cicada endosymbiont Hodgkinia. PNAS. doi: 10.1073/pnas.1421386112.

Cervera-Juanes, R., et al. (2015) MAOA expression predicts vulnerability for alcohol use. Molecular Psychiatry. doi:10.1038/mp.2015.93.

Chwialkowska, K., Nowakowska, U., Mroziewicz, A., Szarejko, I., and Kwasniewski, M. (2016) Water-deficiency conditions differently modulate the methylome of roots and leaves in barley (Hordeum vulgare L.) J. Exp. Bio. doi: 10.1093/jxb/erv552.

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. 

DeWitt, M., Magis, W., Bray, N. et al. (2016, Jan 15). Efficient Correction of the Sickle Mutation in Human Hematopoietic Stem Cells Using a Cas9 Ribonucleoprotein Complex Retrieved from
doi: 10.1101/036236 

Gal, C., et al. (2015) Abo1, a conserved bromodomain AAA‐ATPase, maintains global nucleosome occupancy and organization. EMBO Reports. 16:11. pp 1409 – 1580. doi: 10.15252/embr.201540476.

Gal, C. et. al. (2015) The impact of the HIRA histone chaperone upon global nucleosome architecture. Cell Cycle. 14:1, 123-134. doi:10.4161/15384101.2014.967123.

Gan, H. M., Tan, M. H., Lee, Y. P. and Austin, C. M. (2014) The complete mitogenome of the Australian tadpole shrimp Triops australiensis (Spencer & Hall, 1895) (Crustacea: Branchiopoda: Notostraca).

Gan, H. M., Tan, M. H., Lee, Y. P. and Austin, C. M. (2014) The complete mitogenome of the river blackfish, Gadopsis marmoratus (Richardson, 1848) (Teleostei: Percichthyidae). Mitochondrial DNA. doi:10.3109/19401736.2014.974174.

Givnish, T. J., et al. (2016) Phylogenomics and historical biogeography of the monocot order Liliales: out of Australia and through Antarctica. Cladistics. doi: 10.1111/cla.12153.

Goh, H. F. and Philip, K. (2015) Purification and Characterization of Bacteriocin Produced by Weissella confusa A3 of Dairy Origin. PLoS ONE. doi: 10.1371/journal.pone.0140434.

Gultekin, S. E., et al. (2016) Unusual Presentation of an Adenocarcinoma of the Lung Metastasizing to the Mandible, Including Molecular Analysis and a Review of the Literature. Journal of Oral and Maxillofacial Surgery, doi:10.1016/j.joms.2016.06.004.

Harrisson, K., et al. (2016) Pleistocene divergence across a mountain range and the influence of selection on mitogenome evolution in threatened Australian freshwater cod species. Heredity. 1–10. doi:10.1038/hdy.2016.8.

Hill, C. J. et al. (2016) Effect of room temperature transport vials on DNA quality and phylogenetic composition of faecal microbiota of elderly adults and infants. Microbiome. 4:19. doi: 10.1186/s40168-016-0164-3.

Laver, T., Harrison, J., O’Neill, P. A., Moore, K., Farbos, A., Paszkiewicz, K. and Studholme, D. J. (2015) Assessing the performance of the Oxford Nanopore Technologies MinION. Biomolecular Detection and Quantification 3 (2015) 1–8. doi :10.1016/j.bdq.2015.02.001.

Palomo, A., et al. (2016) Metagenomic analysis of rapid gravity sand filter microbial communities suggests novel physiology of Nitrospira spp. The ISME Journal. doi:10.1038/ismej.2016.63.

Shain, A. H., et al. (2015) The Genetic Evolution of Melanoma from Precursor Lesions. The New England Journal of Medicine. 373:1926-1936. doi: 10.1056/NEJMoa1502583.

Tatarenkova, A., Mesaka F. and Avisea, J. C. (2015) Complete mitochondrial genome of a self-fertilizing fish Kryptolebias marmoratus (Cyprinodontiformes, Rivulidae) from Florida. Mitochondrial DNA. doi: 10.3109/19401736.2015.1115861.

Yang, Y. A., et al. (2016) FOXA1 potentiates lineage-specific enhancer activation through modulating TET1 expression and function. Nucleic Acids Research. doi: 10.1093/nar/gkw498.

Kit Specs

The NEXTflex Rapid DNA-Seq Kit contains enough material to prepare 8 or 48 DNA samples for Illumina® sequencing. The shelf life of all reagents is 12 months when stored properly. All components can be safely stored at -20°C. These kits ship on dry ice.

NEXTflex™ Rapid DNA-Seq Kit Protocol


NEXTflex Rapid DNA-Seq Library Prep Kit for Illumina Sequencing Workflow


Kit Contents

NEXTflex™ End-Repair & Adenylation Buffer Mix
NEXTflex™ End-Repair & Adenylation Enzyme Mix 
NEXTflex™ Ligase Enzyme Mix
NEXTflex™ PCR Master Mix
Nuclease-free Water
NEXTflex™ Sizing Solution
NEXTflex™ Resuspension Buffer


Required Materials Not Provided

1 ng - 1 µg of fragmented DNA in up to 32 µL nuclease-free water.
NEXTflex™ DNA Barcodes – 6 / 12 / 24 / 48 (Cat # 514101, 514102, 514103, 514104) or NEXTflex-96™ DNA Barcodes (Cat # 514106)
Ethanol 100% (room temperature)
Ethanol 80% (room temperature)
96 well PCR Plate Non-skirted (Phenix Research®, Cat # MPS-499) or similar 
96 well Library Storage and Pooling Plate (Fisher Scientific®, Cat # AB-0765) or similar
Adhesive PCR Plate Seal (BioRad, Cat # MSB1001)
Agencourt® AMPure® XP 60 mL (Beckman Coulter Genomics®, Cat # A63880)
Magnetic Stand -96 (Thermo Fisher Scientific®, Cat # AM10027) / or / similar
2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
Nuclease-free barrier pipette tips

Competitive Analysis

The NEXTflex™ Rapid DNA-Seq Kit is compared below with Competitor N’s Kit, illustrating the advantage of Bioo Scientific’s Enhanced Adapter Ligation Technology. The NEXTflex Rapid DNA-Seq Kit and the competitor kit were used to prepare samples through ligation. All samples were then purified and size-selected using an identical AMPureXP bead cleanup step. Triplicate overlays of the three ligation product peaks for NEXTflex Rapid DNA-Seq and Competitor N’s kits were generated. Integration of the ligation product peaks using the Agilent 2100 Bioanalyzer® software produces size-independent values for percentage of product comprising each peak. These values were averaged for the triplicate ligation reactions for each sample. The resulting comparison of unligated, singly-ligated, and doubly-ligated products was graphed. 


Overlays of the triplicate samples of NEXTflex Rapid DNA-Seq and competitor’s kit

Overlays of the triplicate samples of NEXTflex Rapid DNA-Seq (left) and Competitor N’s kit (right).


Enhanced Adapter Ligation Technology

Direct comparison of NEXTflex Rapid DNA-Seq to Competitor N’s kit by analysis of proportions of ligation products. Red portions of the graph indicate non-ligated product proportion; yellow, singly-ligated; green, the desired doubly-ligated amplicon. 


Doubly-ligated product for Illumina DNA library prep

Average triplicate values for the desired doubly-ligated products for NEXTflex Rapid DNA-Seq and Competitor N prepared libraries. The NEXTflex libraries produced 47.1% relative conversion compared to Competitor N’s 21.5%. 

When to Size Select

 Size selection is a critical issue in NGS library preparation. Bioo Scientific’s blog post, Tech Tips – When to Size Select, addresses a number of factors to consider before determining a size selection strategy.


Achieving High Coverage and Yield from GC and AT Rich Genomes


The PCR amplification step in library preparation introduces biases including uneven coverage of regions with extreme base composition, increased numbers of duplicate fragments, decreased mapping quality and poor variant calling. In our new white paper, Achieving High Coverage and Yield from GC and AT Rich Genomes, we show that the NEXTflex™ PCR polymerase contains a highly optimized and robust enzyme that exhibits minimal GC bias and produces uniform coverage of difficult to sequence genomes.


Double SPRI: Obtaining the Proper Insert Length for DNA Library Preparation

Also for a description of a few strategies used to focus read lengths during sample and library preparation read our blog post,Double SPRI: Obtaining the Proper Insert Length for DNA Library Preparation.

DNA Library Prep Whitepapers

White Paper: Optimizing Library Preparation - Enhanced Adapter Ligation Technology

Instructions for using the NEXTflex Rapid DNA-Seq Kit with MYcroarray® Custom Baits for Target Capture

Sequence Capture (hybrid enrichment) – an overview

PerkinEmer Sciclone NGS and NGSx Workstation Automation Guides

Automation Guide for the NEXTflex Rapid DNA-Seq Kit

What tools you should use to analyze your DNA-Seq data?

Read Bioo Scientific’s latest blog post, Analysis Recommendations for DNA-Seq, to find out what tools Bioo Scientific recommends to perform secondary and tertiary analysis.