NEXTflex 16S V4 2.0 (193 -288) 192 RXN

PerkinElmerSKU: PKI-NOVA-4203-06



  • 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


Automated versions of the NEXTFLEX® 16S V4 Amplicon-Seq Library Prep kits 2.0 are available by request.


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® sequencing platforms.


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.



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.


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.


Automate your 16S V4 Library Prep to Increase Throughput and Reduce Errors

A NEXTFLEX® automation protocol is now available on the Sciclone® NGS and NGSx workstations to help labs increase their throughput and reduce human errors. Automated versions of the NEXTFLEX® 16S V4 Amplicon-Seq Library Prep kits 2.0 are available by request.



  • NEXTFLEX® PCR Master Mix
  • NEXTFLEX® 16S V4 PCR I Primer Mix
  • NEXTFLEX® PCR II Barcoded Primer Mix
  • Resuspension Buffer
  • Nuclease-free Water
  • BARCODES 193-288
  • 192 RXNS



  • 1 ng- 50 ng high-quality genomic DNA in up to 36µL 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 1000µL pipettes / multichannel pipettes
  • Nuclease-free barrier pipette tips
  • Vortex
  • 80% Ethanol, freshly prepared (room temperature)



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.



Automation Guide






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