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
For research use only. Not for use in diagnostic procedures.
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.
- 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
- 2, 10, 20, 200 and 1000 µL pipettes / multichannel pipettes
- Nuclease-free barrier pipette tips
- 80% Ethanol, freshly prepared (room temperature)
Selected Citations that Reference the Use of the NEXTFLEX 16S V4 Amplicon-Seq Kit 2.0
- 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.
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.