Description
The NEXTFLEX 16S V3 – V4 Amplicon-Seq Library Prep Kit is designed for the preparation of multiplexed amplicon libraries that span the hypervariable domains three through four (V3 and V4) of microbial 16S ribosomal RNA (rRNA) genes. These libraries are compatible with paired-end sequencing on the Illumina® sequencing platforms.
There are two main steps involved in 16S V3 – V4 amplicon processing: an initial PCR amplification using customized PCR primers that target the V3 and V4 domains, and a subsequent PCR amplification that integrates relevant flow cell binding domains and unique 12 base pair sample indices. The limited number of cleanup steps ensures maximum recovery of amplicons for downstream sequencing.
- Optimized protocol offers lower PCR bias and fewer off-target reads
- Fast library prep protocol
- Low input – As low as 1 ng of genomic DNA
- Flexible barcode options
- Automation protocols are now available for the Revvity Sciclone® NGSx Workstation to automate your 16S sequencing
- Functionally tested on the Illumina® MiSeq® sequencer
Optimized 16S Protocol Offers Lower PCR Bias and Fewer Off-target Reads
The protocol incorporated in the NEXTFLEX 16S V3 – V4 Amplicon-Seq Kit offers better sequencing results than can be obtained using traditional 16S sequencing protocols. The incorporation of the second PCR step in the protocol for the addition of the sample-specific index reduces the number of off-target reads typically encountered during amplicon sequencing.
Increase 16S V3 – V4 throughput and reduce errors with automation
A NEXTFLEX® automation protocol is now available on the Sciclone NGSx workstations to help labs increase their throughput and reduce human errors. Automated versions of the NEXTFLEX 16S V3 – V4 Amplicon-Seq Library Prep kits are available by request.