Illumina NovaSeq X Plus

Published on April 9, 2024

NovaSeq X Plus is a sequencer released by Illumina in January 2023. It is based on Sequencing by Synthesis (SBS).

NovaSeq X Plus is available in the following models:

NovaSeq X Plus - 10B
NovaSeq X Plus - 1.5B
NovaSeq X Plus - 25B

Specifications

Manufacturer	Illumina
Technology	Sequencing by Synthesis (SBS)
Release date	January 2023
Price [2]	$1,250,000 MSRP as of February 2024
Type	Industrial
Weight	532 Kg
Supports real-time analysis	No
Demultiplexing support	Yes
File format	FASTQ
Read length	Illumina NovaSeq X Plus - 10B: 2x50bp to 2x150bp Illumina NovaSeq X Plus - 1.5B: 2x50bp to 2x150bp Illumina NovaSeq X Plus - 25B: 2x50bp to 2x150bp
Run time	Illumina NovaSeq X Plus - 10B: 18 to 25 hours Illumina NovaSeq X Plus - 1.5B: 17 to 23 hours Illumina NovaSeq X Plus - 25B: 48 hours
Reagent cost per Gb [1]	Illumina NovaSeq X Plus - 10B: $3.20 Illumina NovaSeq X Plus - 1.5B: $5.20 Illumina NovaSeq X Plus - 25B: $2.00
Data quality [3]	Illumina NovaSeq X Plus - 10B: ≥ 90% of bases higher than Q30 Illumina NovaSeq X Plus - 1.5B: ≥ 90% of bases higher than Q30 Illumina NovaSeq X Plus - 25B: ≥ 85% of bases higher than Q30
Maximum output per run (Gb)[4]	Illumina NovaSeq X Plus - 10B: 6000.00 Illumina NovaSeq X Plus - 1.5B: 1000.00 Illumina NovaSeq X Plus - 25B: 16000.00
Primary error type [5]	Substitutions

Notes about the specifications

Cost per Gb: The cost per Gb is calculated by dividing the costs of sequencing reagents by the manufacturer’s maximum output per run. For these runs, the manufacturer may have included their own controls and used specific library preparation kits. Actual performance may vary based on sample and library type and quality, loading concentration, and other experimental factors. The sequencing reagent costs used for calculations do not include any other costs associated with preparing the samples or running the instrument, including reagents used for extraction, quality control or library preparation, labor, electricity, lab consumables, storage, etc. Performance metrics and prices are subject to change.
Price: The final price may be different depending on the supplier, the region and any discounts provided through bulk orders or negotiation. Other factors that may influence the final price are warranty options, maintenance contracts, possible trade-ins of older instruments, and any financing arrangements chosen.
Data quality: This is the maximum data quality that the instrument can offer based on manufacturer’s specifications and optimizations. The quality of a specific run will depend on multiple factors, including sample quality, sample quantity, library prep protocol, run conditions, etc.
Maximum output: This is the maximum output of a full instrument run. This number is based on manufacturer’s specifications and optimizations.
Primary error type: This is the most common type of error that may occur due to sequencing. Other types of error may occur as well.

More about the NovaSeq X Plus

The Illumina NovaSeq X Plus is a high-throughput next-generation sequencing (NGS) instrument designed for large-scale, cost-effective genomic research projects. This powerful sequencer targets the high-output segment of the NGS market, making deep sequencing more accessible for projects like large-scale population studies, comprehensive cancer profiling, and high-resolution metagenomics. The NovaSeq X Plus is the clear successor to the popular NovaSeq 6000, offering significantly improved throughput and scalability.

Key Innovations

The NovaSeq X Plus builds upon Illumina's proven sequencing-by-synthesis (SBS) technology but incorporates several advancements to push the boundaries of throughput:

XLEAP-SBS Chemistry: At the heart of the NovaSeq X Plus's performance leap is a novel proprietary chemistry called XLEAP-SBS. This innovative SBS chemistry delivers faster and denser cluster generation on the flow cell. XLEAP-SBS achieves this through a combination of optimized enzymatic components and novel fluorescent nucleotides. The optimized enzymes enhance the speed and efficiency of base incorporation during sequencing cycles. Additionally, the new fluorescent nucleotides boast brighter signal intensity and improved signal separation, allowing for more accurate detection of incorporated bases. Furthermore, XLEAP-SBS employs a two-channel detection scheme to increase sequencing and data processing speed. This innovative approach leverages two fluorescent dyes and two images to determine all four base calls instead of the original four-channel chemistry that uses nucleotides labeled with four different fluorescent dyes.
Increased Flow Cell Capacity: The redesigned flow cell in the NovaSeq X Plus boasts a much higher density of sequencing clusters on the flow cell surface due to reduced spacing between nanowells. Because more clusters can be sequenced simultaneously, the overall data output of the NovaSeq X Plus is dramatically increased, up to 16 Tb with the 25X flow cell.
Enhanced Onboard Computing: The NovaSeq X Plus integrates powerful computational resources directly into the instrument itself. This on-board cluster facilitates much faster primary data analysis (base calling and demultiplexing) compared to traditional sequencing workflows that rely on transferring large datasets to external servers for processing. This significantly reduces turnaround times for initial data analysis, expediting downstream analyses and accelerating project timelines. In some cases, secondary analysis may even be possible through an onboard DRAGEN pipeline, enabling researchers to analyze data on the instrument right after sequencing.

Features and Technologies

Ultra-High Throughput: The NovaSeq X Plus can generate an astonishing 16 Tb of data in a single run using with the dual flow cell feature.
Scalability: The dual flow cell system enables flexible output based on project needs.
Accuracy: It employs proven Illumina SBS chemistry, delivering highly accurate sequencing data with industry-leading low error rates.
DRAGEN for Onboard Analysis: Integrated analysis software enables secondary data analysis directly on the instrument.

Intended Uses and Specific Applications

The NovaSeq X Plus is designed for applications requiring extremely high sequencing output:

Whole Genome Sequencing (WGS): Population genomics, large-cohort studies, or disease research (trios). Compatible library prep kits include Illumina DNA PCR-Free and Illumina TruSeq DNA Nano.
Whole Exome Sequencing (WES): For identifying coding region variants. A compatible kit is the Illumina TruSeq DNA Exome kit.
RNA Sequencing: Comprehensive transcriptome profiling, gene expression analysis, and identification of novel transcripts. Compatible kits include Illumina Stranded Total RNA Prep with Ribo-Zero Plus, Illumina Stranded mRNA Prep.
Metagenomics: Deep sequencing of complex microbial communities. Compatible library prep kits include Illumina Nextera DNA Flex Library Prep Kit and Illumina DNA PCR-Free Library Prep Kit.

Comparison with Competing Instruments

Illumina NovaSeq 6000: The NovaSeq X Plus significantly surpasses the NovaSeq 6000 in output per run (~60% more), while maintaining comparable cost per gigabase. This makes the X Plus the optimal choice for projects demanding maximum throughput.
Complete Genomics DNBSEQ-T7: The DNBSEQ-T7 run time is faster than the 25X flow cell of the NovaSeq X Plus. However, the NovaSeq X Plus maintains a significant throughput advantage.
Complete Genomics DNBSEQ-T20x2: This system is capable of higher output than the NovaSeq X Plus but may have higher upfront costs and longer run times in some configurations.

Special Considerations

The NovaSeq X Plus is an excellent choice if:

Maximum data output per run is essential for your project.
Your research involves extremely large sample numbers or highly complex genomes/transcriptomes.
Cost-effectiveness at scale is a concern.

Need sequencing done on a Illumina NovaSeq X Plus or a specific instrument? Tell us about your project here, and we'll connect you with a qualified provider.

This information on this page is up-to-date as of April 9, 2024 and based on the spec sheets published by the manufacturer.