16S metagenomic sequencing enables targeted exploration of microbial communities by combining amplification of the 16S ribosomal RNA (rRNA) gene, a reliable phylogenetic marker, with next generation sequencing (NGS) technology.
INTEGRA Biosciences has created an application guide that ensures precise amplicon sequencing, allowing the study of a wide variety of microbial communities.
Custom primers are used to amplify variable regions three and four (V3 and V4) of the 16S rRNA gene, which are then combined with Illumina sequencing adapters and dual indices for multiplexing.
This method, which uses paired-end 300 bp reads, produces high-quality data for taxonomic classification at the genus and species level.
INTEGRA provides ASSIST PLUS programs for each of these processes, which can be further optimized for your workflows, enabling straightforward, automated 16S metagenomic sequencing library creation.
Key benefits
Optimized throughput: Automates up to 24 samples per run, improving efficiency in medium-throughput facilities.
Reduced time and effort: Simplifies complex operations to improve productivity.
Convenient walk-away processing: Four easy-to-adapt VIALAB template scripts reduce user intervention and manual handling for hands-free operation.
Automated bead handling: Standardized bead clean-up with the integrated MAG module improves precision, reduces mistakes, and improves reproducibility.
Overview: How to perform automated 16S rRNA library preparation
The ASSIST PLUS pipetting robot simplifies 16S metagenomic sequencing library preparation while increasing throughput, precision, and dependability.
Four preprogrammed VIALAB template scripts efficiently handle the key steps of the 16S library preparation procedure for up to 24 samples.
INTEGRA also provides verified algorithms for normalizing and pooling in other app notes. These automated, walk-away methods will improve user convenience and standardization, resulting in greater reproducibility of results and reduced hands-on time.
Elevate your research with the ASSIST PLUS: get consistent and accurate results while saving time.
The bead clean-up stage is a vital part of 16S library preparation, and can be readily completed using an INTEGRA MAG module. This magnetic separation system automates magnetic bead handling by moving vertical magnet arrays, eliminating the need for unwanted manual interventions.
Image Credit: INTEGRA Biosciences
Conclusion
This application guide provides free VIALAB templates for 16S library preparation, including amplification and clean-up, which may be used to construct automated protocols.
Detailed reagent and deck layout guidance reduces set-up errors and improves reproducibility, promoting technique uniformity in different lab conditions.
The ASSIST PLUS, when combined with specialized modules like the MAG and COLDPLATE, allows efficient magnetic bead separation and precise thermal control, supporting critical steps in PCR and purification.
The automated workflow on the ASSIST PLUS is modular, allowing easy customization and scalability to adapt to future protocol updates or changing sample input requirements.
About INTEGRA Biosciences
INTEGRA provides innovative solutions for liquid handling and media preparation applications that serve the needs of its customers in research, diagnostics and quality control laboratories.
The company’s instruments and plastic consumables are developed and manufactured in Zizers, Switzerland and Hudson, NH USA. In order to remain close to its customers, the company maintains a direct sales and support organization in North America, the UK, France and Germany, as well as a network of over 100 highly trained distribution partners worldwide.
In recent years INTEGRA has focused on developing a new and technologically advanced range of handheld electronic pipettes which are simple to use and meet the ergonomic needs of their customers. Today, the company is proud to offer the widest range of electronic pipettes in the market spanning a range from single channel pipettes up to 384 channel benchtop instruments.
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