xGen™ microbial amplicon panels

The 16S v2 and ITS1 NGS panels are designed to study complex microbial communities using a single set of primers targeting the 16S rRNA gene or ITS1 region.

More detailed product information

The 16S v2 and ITS1 NGS panels are designed to study complex microbial communities using a single set of primers targeting the 16S rRNA gene or the ITS1 region. Additional targets can be added to these panels if required.

xGen 16S rRNA v2 is a panel targeting the V1-V9 variable regions of the 16S rRNA gene
xGen ITS1 focuses on the ITS (internal transcribed spacer regions) of rRNA genes
Simple, two-hour workflow, in one tube
Integrated library normalization enables simplified library balancing and pooling without the need for sample quantification
Unique amplicon chemistry that generates high-diversity clusters even without PhiX, ensuring more efficient sequencing

Features	Specification
Panel information	16S v2: 23 primers, average amplicon size 425 bp ITS1: 15 primers, amplicon size 145–695 bp
Input material	10 pg for microbial isolates; 1-50 ng for metagenomic samples
Time	2 hours from cDNA to library or 3 hours from cDNA to normalised library pool
Multiplexing	Up to 96 combinatorial dual index (CDI) or 1536 unique dual index (UDI)
Recommended reads	16S v2: 100K reads per library ITS1: 25K reads per library

xGen Amplicon Panels have a single tube workflow that completes in 2 hours. Creating an NGS library starts with multiplex PCR. Your panel is combined with the DNA sample to amplify the targets of interest. The samples are then amplified with indexing primers to create a functional dual indexed library. As an optional step, the xGen Normalase reagent can be used after pooling multiple libraries to ensure each is equally represented in the final sample for the flowcell.

Primer coverage of xGen 16S Amplicon Panel v2

xGen 16S Amplicon Panel v2 (primers). Sequencing read coverage for an E. coli DNA sample (n = 1) observed in Integrative Genomics Viewer (IGV) Sashimi plot and illustration of multiplexed primer coverage of all nine variable regions of 16S rRNA compared to a standard V3/V4 16S sequencing read coverage.

Primer coverage of xGen ITS1 Amplicon Panel

xGen ITS1 Amplicon Panel (primers). Sequencing read coverage observed in the IGV Sashimi plot and illustration of multiplexed primer coverage of ITS1 region for Candida albicans (n = 1). Reads originating from the forward primer are shown in red; reads from the reverse primer are in blue.

Representation of microbial communities in xGen 16S Amplicon Panel

The xGen 16S Amplicon Panel v2 provides superior representation of diverse microbial communities. The 16S v2 panel covering V1–V9 regions of 16S rRNA provides accurate representation of each genus in a commercially available standard (MSA-1003) compared to libraries interrogating the V3–V4 region alone. The legacy Accel-Amplicon 16S+ITS panel and the new xGen 16S Amplicon Panel v2 show similar relative abundance results. Strains were present at levels from 0.02% to 18% in MSA-1003. Boxed organisms were not detected with sole use of the V3-V4 region. Libraries were sequenced on a MiSeq^® (Illumina) instrument without the use of the PhiX reagent. As shown in the examples in this figure, by not losing reads to PhiX or having to sequence deeper due to the use of phased primers, an increased number of samples can be fit on a sequencing run, thereby increasing sequencing efficiency, while still achieving quality data.

For more information, please contact us by e-mail: diagnostika@pentagen.cz