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

Validated library prep for 16S V4 rRNA sequencing using Illumina® sequencing primers

 

Catalog# Product Name Quantity US List Price
4203-01
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (4 Barcodes)
8 rxns $180 Buy Now
4203-02
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (12 Barcodes) 24 rxns $423 Buy Now
4203-03
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (48 Barcodes) 96 rxns $1,524 Buy Now
4203-04
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (Barcodes 1- 96)
192 rxns $2,895 Buy Now
4203-05
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (Barcodes 97 - 192)
192 rxns $2,895 Buy Now
4203-06
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (Barcodes 193 - 288)
192 rxns $2,895 Buy Now
4203-07
NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 (Barcodes 289 - 384) 192 rxns
$2,895 Buy Now

NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0
  • 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 Workstation to automate your 16S sequencing
  • Functionally validated with Illumina® sequencing platforms

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 and are ideal for monitoring population fluctuations of microbial communities.

Genera from canine feces measured by analysis of 16S V4 reads

Genera measured by analysis of 16S V4 reads obtained from library constructed with the NEXTflex 16S V4 Amplicon-Seq Kit 2.0 from 20 ng of DNA isolated from canine feces.


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 Specs

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.

NEXTflex 16S V4 Amplicon-Seq Kit 2.0 Protocol

 

NEXTflex™ 16S V4 Amplicon-Seq Kit 2.0 Protocol

 

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 of NEXTflex 16S V4 Amplicon-Seq 2.0 Barcoded Primers Indexes - Excel / PDF

Instructions for installing NEXTflex Barcode Indices in Illumina® Experiment Manager