NEXTFLEX 16S V4 Amplicon-Seq Kit 2.02018-10-18T03:59:20+00:00

VALIDATED 16S V4 rRNA LIBRARY PREP FOR BROAD-RANGE ANALYSIS OF MIXED MICROBIAL POPULATIONS

NEXTFLEX® 16S V4 Amplicon-Seq Kit 2.0 for Illumina® Platforms

  • Fast and easy library prep protocol
  • Low input – Only 1 ng of input required
  • Multiplex up to 384 libraries
  • Custom sequencing primers are not required
  • Automation protocols are now available for the Sciclone® NGS and NGSx Workstations to automate your 16S sequencing
  • Functionally validated with Illumina® sequencing platforms
Download Manual
Download Sciclone Automation Guide
For research use only. Not for use in diagnostic procedures.
AUTOMATED

  • NOVA-4203-01

       $186

    4 BARCODES
    8 RXNS
AUTOMATED

  • NOVA-4203-02

       $436

    12 BARCODES
    24 RXNS
AUTOMATED

  • NOVA-4203-03

       $1,570

    48 BARCODES
    96 RXNS
AUTOMATED

  • NOVA-4203-04

       $2,981

    BARCODES 1-96
    192 RXNS
AUTOMATED

  • NOVA-4203-05

       $2,981

    BARCODES 97-192
    192 RXNS
AUTOMATED

  • NOVA-4203-06

       $2,981

    BARCODES 193-288
    192 RXNS
AUTOMATED

  • NOVA-4203-07

       $2,981

    BARCODES 289-384
    192 RXNS

Ideal for Monitoring Populations of Microbial Communities

The NEXTFLEX® 16S V4 Amplicon-Seq Library Prep Kit 2.0 is designed for the preparation of multiplexed amplicon libraries spanning the V4 hypervariable domain of microbial 16S ribosomal RNA (rRNA) genes. These libraries are compatible with paired-end sequencing on the Illumina® MiSeq® sequencing platform.

Optimized Protocol Offers Lower PCR Bias and Fewer Off-target Reads

The NEXTFLEX 16S V4 Amplicon-Seq Kit 2.0 incorporates a second PCR step in the protocol for the addition of the sample-specific index which reduces off-target reads encountered during amplicon sequencing.

Low Input Requirements for 16S rRNA Sequencing

As little as 1 ng of genomic DNA can be used to generate libraries using the NEXTflex 16S V4 Amplicon-Seq Library Prep Kit 2.0.

384 Barcodes Available for Cost-effective 16S V4 rRNA Sequencing

The NEXTFLEX 16S V4 Amplicon-Seq Library Prep Kit 2.0 can be used for multiplexing up to 384 samples, to greatly reduce the cost of sequencing. The barcodes are offered in sets of 4, 12, 48 or 96 barcodes and supplied with 2 reactions worth of each barcode.

KIT CONTENTS

  • NEXTFLEX® PCR Master Mix
  • NEXTFLEX® 16S V4 PCR I Primer Mix
  • NEXTFLEX® PCR II Barcoded Primer Mix
  • Resuspension Buffer
  • Nuclease-free Water

REQUIRED MATERIALS NOT PROVIDED

  • 1 ng – 50 ng high-quality genomic DNA in up to 36 µL nuclease-free water for each library
  • 96 well PCR Plate Non-skirted (Phenix Research, Cat # MPS-499) or similar
  • Adhesive PCR Plate Seal (Bio-Rad®, Cat # MSB1001)
  • Agencourt® AMPure® XP 5 mL (Beckman Coulter® Genomics, Cat # A63880)
  • Magnetic Stand – 96 (Thermo Fisher® Scientific, Cat # AM10027) or similar
  • Thermocycler
  • 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
  • Nuclease-free barrier pipette tips
  • Vortex
  • 80% Ethanol, freshly prepared (room temperature)

SEQUENCES

  • Sequences of NEXTFLEX 16S V4 Amplicon-Seq 2.0 Barcoded Primers Indexes – Excel / PDF

Selected Citations that Reference the Use of the NEXTFLEX 16S V4 Amplicon-Seq Kit 2.0

  • Bruce-Keller, A. J., Fernandez-Kim, S., Townsend, R. L., Kruger, C., Carmouche, R., Newman, S., . . . Berthoud, H. (2017). Maternal obese-type gut microbiota differentially impact cognition, anxiety and compulsive behavior in male and female offspring in mice. Plos One, 12(4). doi:10.1371/journal.pone.0175577.
  • Luk, B., Veeraragavan, S., Engevik, M., Balderas, M., Major, A., Runge, J., . . . Versalovic, J. (2018). Postnatal colonization with human “infant-type” Bifidobacterium species alters behavior of adult gnotobiotic mice. Plos One, 13(5). doi:10.1371/journal.pone.0196510.
  • Luna, R. A., Oezguen, N., Balderas, M., Venkatachalam, A., Runge, J. K., Versalovic, J., . . . Williams, K. C. (2017). Distinct Microbiome-Neuroimmune Signatures Correlate With Functional Abdominal Pain in Children With Autism Spectrum Disorder. Cellular and Molecular Gastroenterology and Hepatology, 3(2), 218-230. doi:10.1016/j.jcmgh.2016.11.008
  • Moon, C., Stupp, G. S., Su, A. I., & Wolan, D. W. (2017). Metaproteomics of colonic microbiota unveils discrete protein functions among colitic mice and control groups. doi:10.1101/219782.
  • Stalenhoef, J. E., et al. (2017) Fecal Microbiota Transfer for Multidrug-Resistant Gram-Negatives: A Clinical Success Combined With Microbiological Failure. Open Forum Infectious Diseases, 4(2). doi:10.1093/ofid/ofx047.
  • Svensson, K., Paruch, L., Gaby, J. C., & Linjordet, R. (2018). Feeding frequency influences process performance and microbial community composition in anaerobic digesters treating steam exploded food waste. Bioresource Technology, 269, 276-284. doi:10.1016/j.biortech.2018.08.096.
  • Sylvia, K. E., Deyoe, J. E., & Demas, G. E. (2018). Early-life sickness may predispose Siberian hamsters to behavioral changes following alterations of the gut microbiome in adulthood. Brain, Behavior, and Immunity, 73, 571-583. doi:10.1016/j.bbi.2018.07.001.
  • Whon, T. W., Chung, W., Lim, M. Y., Song, E., Kim, P. S., Hyun, D., . . . Nam, Y. (2018). The effects of sequencing platforms on phylogenetic resolution in 16 S rRNA gene profiling of human feces. Scientific Data, 5, 180068. doi:10.1038/sdata.2018.68.
  • Wotzkas, S. Y., Kreuzer, M., Maier, L., Zuend, M., Schlumberger, M., Nguyen, B., . . . Misselwitz, B. (2018). Microbiota stability in healthy individuals after single-dose lactulose challenge – a randomized controlled study. doi:10.1101/424531.
  • Zhao, R., Yang, W., Pei, F., Zhao, L., & Hu, Q. (2018). In vitro fermentation of six kinds of edible mushrooms and its effects on fecal microbiota composition. Lwt, 96, 627-635. doi:10.1016/j.lwt.2018.06.012.

The NEXTFLEX 16S V4 Amplicon-Seq Kit 2.0 contains enough material to prepare 8, 24, 96 or 192 amplicon-seq libraries from genomic DNA for Illumina® platforms. The shelf life of all reagents is 12 months when stored properly. All components can be safely stored at -20°C. This kit is shipped on dry ice.

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