NGS solution for library preparation aimed at the simultaneous identification and profiling of microbial species present in DNA extracted from original samples, through the analysis of the variable regions V1-V4 of the 16S rRNA gene. Product for research use only (RUO) and not intended for diagnostic procedures.
Detailed Description
Principle of operation
VIASURE 16S V1-V4 NGS Solution is a non‑automated device for NGS library preparation based on amplicon enrichment. The assay amplifies target regions of the bacterial 16S rRNA ribosomal gene, specifically the variable domains V1 to V4, and simultaneously adds unique barcodes and indexes necessary for subsequent sequencing.
The workflow is based on a single qPCR reaction that combines the enrichment of target sequences and sample indexing using specifically designed primers. After this reaction, individual libraries are pooled by equal volume or relative quantification, a single final library purification is performed, quantified, diluted, and prepared for sequencing.
Relative quantification of individual libraries is possible thanks to the final fluorescence signal generated by a DNA binding dye included in the reaction mix; this signal is proportional to the amount of target template and can be measured on real‑time PCR platforms.
Applications
The product is designed to facilitate the identification and profiling of microbial species present in DNA extracted from research samples, by analyzing variable regions of the 16S rRNA gene. 16S rRNA gene sequencing is described as a widely used technique to study the composition and diversity of the microbiome in microbial communities.
Commercial literature indicates that the product supports biological samples or cultures and is aimed at obtaining reliable and accurate results in NGS‑based research applications.
Benefits
Allows coverage of the V1, V2, V3, and V4 regions of the 16S rRNA gene using a OneStep multiplex qPCR strategy.
Integrates target region amplification and barcode incorporation for sequencing into a single qPCR.
Reduces workflow complexity through a simplified process of qPCR, quantification, pooling, and purification.
qPCR allows immediate and accurate quantification of libraries, according to the product leaflet.
Requires only a single final library purification after pooling.
Includes access to Certest bioinformatics software for sequencing data analysis in FASTQ format and generation of Krona reports.
Key results or indicators
Library size: 500-600 bp.
DNA input requirement: 5 µl of sample.
Number of pools per panel: 1.
Plexity: up to 48.
Hands‑on time: <30 min.
Platforms: Illumina MiSeq and Element Biosciences AVITI.
Technology used
The assay uses an amplicon‑based enrichment NGS library preparation strategy, compatible with Illumina sequencing platforms. The library is generated from isolated microbial DNA, by qPCR for V1-V4 region amplification and sample indexing.
The workflow includes:
Enrichment and indexing.
Pooling.
Final library purification.
Sequencing.
Data analysis.
Analysis software
The solution includes access to Certest bioinformatics software for raw sequencing data analysis (V-Xplora). It is a web application developed by Certest Biotec S.L. that allows generating Krona files or reports from FASTQ files.
Intended user / audience
The product is intended for qualified laboratory personnel trained in Next Generation Sequencing (NGS) techniques, nucleic acid extraction, sequencer handling, and analysis using pipelines or bioinformatics software.
Considerations or limitations
Product for research use only (RUO).
Not intended for diagnostic procedures.
No declared clinical purpose and should not be used for diagnosis, prevention, or treatment of diseases.
Product performance characteristics have not been established for diagnostic use.
Some species are better classified by certain variable regions than others; the user must know the regions of interest for classification during bioinformatic analysis.
Because the 16S rRNA gene is highly conserved and widespread among bacteria, environmental contamination may occur if precautions are not strictly followed during the procedure.
Key Features
NGS solution for 16S rRNA gene analysis.
Coverage of variable regions V1, V2, V3, and V4.
Library preparation using OneStep multiplex qPCR.
Enrichment and indexing in a single reaction.
Amplicon‑based method.
Compatible with Illumina MiSeq and, according to the leaflet, Element Biosciences AVITI.
Supports biological samples or cultures.
Library size of 500-600 bp.
Up to 48 samples.
Hands‑on time <30 min.
Single pool per panel.
Data analysis using V-Xplora / VIASURE in‑house software.
RUO product, not intended for diagnosis.
Presentation Details
Format and reference
Product: VIASURE 16S V1-V4 NGS Solution
Reference: VS-16S-V1448PHRUO
Use: Research Use Only (RUO)
Number of reactions: 48
Product type: non‑automated solution for NGS library preparation
Supplied content
MIX 1 16S V14: lyophilized master mix containing primers, DNA binding dye, and components necessary for the generation, enrichment, and indexing of libraries of the target regions. Presentation: 1 plate with 48 wells.
MIX 1 Rehydration buffer: solution to reconstitute the stabilized MIX 1 product. Presentation: 1 vial of 1.8 ml.
Tear-off-8-cap strips: optical caps for sealing during thermal cycling. Presentation: 1 bag with 6 strips of 8 caps.
Transport, storage and handling conditions
The product can be transported and stored between 15-30 °C until the expiration date indicated on the label.
Vibrations during transport must be avoided to prevent leaks.
It is recommended not to rehydrate wells that will not be used.
Unused wells must be kept in the supplied aluminum bags with silica gel, protected from light.
Once wells are rehydrated, nucleic acid must be added and PCR performed immediately.
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