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Novogene’s Revio Data: Achieving Maximum 110Gb HiFi Reads per SMRT Cell!

Novogene has recently installed PacBio’s latest high-throughput sequencing system—Revio. Revio delivers exceptional performance in terms of throughput with excellent base quality. This cutting-edge technology promises a historically high throughput of 360 Gb of HiFi reads per day, making a significant 15× increase compared to the previous Sequel II system. Novogene has validated runs with DNA HiFi libraries of plants, animals and human. The results show that the system may produce a maximum output of up to 110.5 Gb per SMRT cell, which surpass the standard data output of 90Gb per SMRT cell published by PacBio. The enhanced capabilities of Revio system will allow Novogene to add high throughput to HiFi long reads sequencing and direct methylation detection for a wide array of applications.

1.HiFi yield per SMRT cell of Revio system

The average data output per SMRT cell is 98.4Gb, with a quality value of 31.2. Additionally, the platform demonstrates a maximum data output of 110.5 Gb per SMRT cell.

Table 1 Quality control result of Revio sequencing data

Sample HiFi Yield (Gb) Read Length(mean)
(bp)
Read Length(N50)
(bp)
Mean Quality Value
Human Sample 1 110.5 21,313 21,967 31.3
Animal Sample 1 98.3 16,305 16,486 31.1
Plant Sample 1 86.4 19,659 19,953 31.1
2.Testing results of multiplexed libraries

DNA multiplexed libraries, consisting of three animal samples and three plant samples are sequenced on Revio system respectively. The results show that each animal sample yields more than 30Gb HiFi reads, and the data demultiplexing rate reaches a level of more than 99%.

Table 2 Quality control result of multiplexed SMRT cell libraries

Sample Total HiFi Yield (Gb) Data Demultiplexing Rate (%) Single-sample HiFi Yield (Gb) Read Length(mean)
(bp)
Read Length(N50)
(bp)
Animal 1 102.3 99.56 32.0 20,634 20,558
Animal 2 34.2 19,523 19,464
Animal 3 35.7 19,074 18,988
Plant 1 84.6 99.76 30.0 17,157 17,002
Plant 2 23.1 16,634 16,614
Plant 3 31.2 16,513 16,320
3.Comparison of sequencing data achieved on Sequel II system and Revio

Human and plant gDNA samples are used for HiFi library preparation and sequenced on PacBio’s Sequel II system and Revio system respectively to evaluate the performance differences. The results indicate that the Revio system is able to yield 2-3 times more HiFi data per SMRT cell compared to Sequel II. Furthermore, Revio completes the sequencing in a shorter run time, while maintaining a mean data quality > Q30.

Table 3 Sequel II system vs Revio system sequencing data

Sample System HiFi Yield (Gb) Read length(mean)
(bp)
Read length(N50)
(bp)
Run times
(h)
Mean quality value
Human Sample 1 Sequel II 38.4 20,420 20,345 30 31.0
Human Sample 1 Revio 110.5 21,313 21,967 24 31.3
Plant Sample 1 Sequel II 31.8 15,462 15,555 30 31.1
Plant Sample 1 Revio 84.6 18,799 19,338 24 31.1
4.Data analysis of human whole genome sequencing on Revio system

The achieved mapping rate of human whole genome is 99.91%, with coverage across the entire genome of 99.64% and an average sequencing depth of 38X.

Table 4 Mapping statistics of human samples sequenced on the Revio system.

Species HiFi Yield (Gb) Clean Reads Mapped Reads Mapping Rate (%) 1X Coverage Rate (%) 4X Coverage Rate (%) 10X Coverage Rate (%) 20X Coverage Rate (%) Mean Depth
Human Sample 110.5G 5,555,691 5,550,504.00 99.91 99.64 99.26 98.67 94.37 38
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b

Figure 1 Analysis results of human whole genome resequencing (WGS) data

Figure 1. (a): Statistics of structural variation length. (b): The circos figure shows the density of variant chromosome distribution, including translocation, insertion, deletion, inversion, copy number variation, short tandem repeat, gene, chromosome.

About Novogene

Novogene is a pioneer in applying cutting-edge molecular biology technology and high-performance computing to research in the fields of life science and human health. Our vision is to be the global leader in providing genomic services and solutions.

Novogene has extensive coverage in global genome projects and has accumulated vast experiences in third-generation library preparation, sequencing, and bioinformatics analysis for numerous species. Novogene has technical advantages in de novo sequencing technology and applications, holding 18 patent licenses and over 40 software copyrights. Our vision is to be the global leader in providing genomic services and solutions.

With one of the largest sequencing capacities in the world, we utilize our deep scientific knowledge, first-class customer service, and unsurpassed data quality to help clients realize their research goals in the rapidly evolving world of genomics. Novogene is committed to becoming your trusted genomics partner.