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Revio 系统 + SPRQ 化学

精准的长度长测序在一定程度上为您揭示更多信息

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HiFi sequencing at scale

With a high-density SMRT Cell, up to 4 SMRT Cells per run, and 24-hour run times3, the Revio system with SPRQ chemistry delivers up to 480 Gb2,5 of HiFi reads per day, equivalent to 2,500 human whole genomes4 per year. ​

The $5006 complete, phased genome

HiFi sequencing provides small variants, structural variants, repeat expansions, methylation, and haplotype phasing from a single library and sequencing run. With a comprehensive genome, you can replace multiple assays, saving valuable time and resources while gaining deeper insights in one streamlined process.

Long-read genomes at scale, with 4x less DNA input required

SPRQ chemistry on the Revio system unlocks the ability to sequence more sample types than ever before, using just 500 ng of native DNA – without sacrificing the high throughput or exceptional quality you rely on.

On-instrument 5mC and 6mA caller for multiomic Fiber-seq chromatin assay

SPRQ chemistry, paired with Google Health DeepConsensus algorithms, delivers exceptional read accuracy plus confident 5mC and 6mA methylation detection in every run. Optimized file formats reduce data storage needs through quality value binning and smart read ordering, streamlining data handling and maximizing efficiency.

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Image of Revio system facing left - PacBio

简化的耗材

Revio 系统用的耗材更少，优化后便于追踪且大大减少浪费，从而简化了 HiFi 测序体验。工作流程改进后，花不到一分钟时间就能将耗材加载到仪器上。

Revio SPRQ 聚合酶试剂盒

Optimized reagents to bind polymerase to prepare 24 SMRTbell libraries for sequencing​

Revio SMRT Cell 托盘

一个托盘包含 4 张高密度 SMRT Cell 芯片，每张都有 2500 万个零模波导孔。

Revio SPRQ 测序板

Reagents for sequencing up to 4 Revio SMRT Cells. The sequencing plate combines multiple parts from the Sequel IIe system: sequencing plate, tube septa, sample plate, mixing plate, and SMRT Cell oil.

HIFI LEVEL ACCURACY AND DATA QUALITY

 

The Revio system with SPRQ chemistry produces exceptional read length, quality, and variant calling performance that have made HiFi sequencing so celebrated.

Reveal a more complete genome​

The complete, gapless human genome marks a new era of genomics where no region of the genome is beyond your reach. In 2021, PacBio HiFi technology was used by the Telomere-to- Telomere (T2T) Consortium to fill in those gaps.

Read length

HiFi reads are tens of kilobases long, providing the ability to resolve large variants and map to difficult regions of the genome.​

Read and base quality

The Revio system provides reliable answers through exceptional accuracy, with 90% of bases Q30+ and median read accuracy ≥Q30.​

Variant calling

HiFi sequencing with SPRQ chemistry on the Revio system delivers high accuracy variant detection, resulting in fewer sequencing errors relative to other technologies at lower coverage1.

强大的计算能力

Revio 系统内置多个强大的 NVIDIA GPU，提供的计算能力超出 Sequel IIe 系统 20 倍，通过仪器即可直接输出即时可用的准确数据。

HiFi read 生成

GPU 可为碱基检出与 HiFi read 生成缩短周转时间，直接以 BAM 格式输出 HiFi reads。

Google Health DeepConsensus

DeepConsensus 使用高级深度学习技术进一步提高了 HiFi 测序的准确度与产量。

甲基化检出

标准运行包括 DNA 甲基化状态，使用能处理聚合酶动力学的深度学习算法计算。

拆分

包括条形码拆分在内的所有基本处理步骤均在仪器上完成。

 

“…Revio 系统的高通量功能让我们能把 HiFi 技术应用到农业生物科技领域的大量应用中“

 ——Corteva Agriscience 生物技术与基因组学技术经理 Gina Zastrow-Hayes

REVIO 长读长测序方法对比

PacBio Revio with SPRQ SBS 测序 纳米孔测序
读长 15-20 kb5 2×150 bp 10-100 kb
Read 准确度 99.95% (Q33)1 99.92% (Q31)3 98.90% (Q19)4
运行时间 24 小时2 44 小时 72 小时
Yield 120-480 Gb2 2,400–3,000 Gb 50–110 Gb
Coverage Unbiased Reduced at low and high (GC) Reduced in low-complexity runs
Genome completeness6
✓
X
✓
Variant calling - SNVs, Indels, SVs5
✓
X
X
Areas of high homology
✓
X
X
Phasing
✓
X
✓
Methylation - 5mC and 6mA Every run Via library prep Every run

1. Read accuracy PacBio HiFi: precisionFDA Truth Challenge V2​
2. HiFi yield specification based on HG002/GM24385 human DNA extracted with Nanobind CBB kit and prepared with SMRTbell prep kit 3.0. Run time specification is for the sequencing reaction. HiFi yield is dependent on library fragment size. Yield is typically lower for shorter libraries.​

3. Illumina: HG002 2×150 bp NovaSeq library, precisionFDA Truth Challenge V2​
4. ONT: Q20+ chemistry (R10.4, Kit 12), Oct 2021 GM24385 Q20+ Simplex dataset release​
5. Shafin Nat Methods 2021 (https://doi.org/10.1038/s41592-021-01299-w)​
6. Cheng 2021 Nat Methods (https://doi.org/10.1038/s41592-020-01056-5) and Shafin 2020 Nat Biotech (https://doi.org/10.1038/s41587-020-0503-6)

 

工作流程

提取

Nanobind DNA 试剂盒可从各种样本类型中提取 HMW gDNA。

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文库制备

SMRTbell 制备试剂盒 3.0 支持人工与自动文库制备，能轻松创建 SMRTbell 文库。

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测序

Revio 系统能结合自身时间表灵活安排运行时间，因此可连续 24 小时同步运行 4 个 SMRT Cell。

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数据分析

Google Health 的高级算法与前沿的 NVIDIA GPU 可实现机载测序数据处理，输出文件大小均有缩小。

了解更多

更多 HIFI READS，更多样本，更多发现

System specifications with SPRQ chemistry

The Revio system utilizes nanofabricated Revio SMRT Cells and has four independent stages, allowing sequencing of multiple SMRT Cells in parallel. The onboard compute provides accurate basecalling with Google DeepConsensus, plus methylation calling in every run.

Library Run timea Q30+ bases HiFi yield per SMRT Cellb Methylation
1-5 kb 12 hours 95% 6-8 million reads 5mC at CpG sites and 6mA for native DNAc
5-10 kb 24 hours 90% 35-70 Gb
10-15 kb 70-100 Gb
15-20 kb 100-120 Gb
20-25 kb 30 hours 85% 100-120 Gb
  1. Run time refers to the data collection step, which determines the time between processing SMRT Cells.
  2. HiFi yield is dependent on library quality and sequencing preparation procedures.  Specified yield is based on high-quality samples prepared following best practices. 
  3. The Revio 6mA caller is designed to detect methylation in the context of the Fiber-seq chromatin assay.

 

仪器规格

运行环境
温度 19–25°C (66–77°F)
湿度 20%–80% 相对湿度，无冷凝
通风 13,000 BTU/h (3,800 W)
尺寸
W × D × H 92.7 厘米（36.5 英寸） × 91.4 厘米（36.0 英寸）× 174.5 厘米（68.7 英寸）
重量 465 千克（1,025 磅）
占地面积 243 厘米（96.0 英寸）× 138.5 厘米（54.5 英寸）
电功率
所需电源 200–240 VAC，50-60 Hz，max 5 kVA/4.8kW/24A
计算
网络连接 1 GbE 或 10 GbE 铜, RJ45 金属连接头
输出文件格式 hifi_reads.bam，每个文件大小 ≈55 Gb，每年高达 72 TB

下载规格表     访问数据集

常见问题

The SPRQ (pronounced “spark”) chemistry is our newest HiFi chemistry that makes sequencing more efficient and expands HiFi sequencing to more sample types and applications. SPRQ requires only 500 ng of native DNA, which opens up even more possibilities for your projects. Plus, SPRQ helps lower genome costs by increasing HiFi yields per SMRT Cell by 33%, allowing you to sequence two human genomes at 20X coverage per SMRT Cell. And that’s not all—SPRQ also enables deeper multiomic insights into chromatin architecture with Fiber-seq, using on-instrument 5mC and 6mA calling for greater precision in methylation detection.

The Revio system with SPRQ chemistry sequences two human genomes at 20x coverage for just $5006. Your local sales representative can provide detailed pricing in your currency. 6$500 per human HiFi genome assumes US list price of $995 for sequencing reagents for one Revio SPRQ SMRT Cell, which has an expected yield of 120 Gb, equivalent to two 20× human genomes.

单个 Revio SMRT Cell 所需测序试剂在美国的市场价为 995 美元，预期产量为 90 Gb，相当于 30X 人类基因组。您当地的销售代表可提供以当地货币表示的价格详情

是。同样的文库制备协议对 Sequel 系统与 Revio 系统均有效。因此，SMRTbell 文库可以跨系统使用。不过，各个系统使用的测序试剂（聚合酶/结合试剂盒、测序盘与 SMRT Cell 托盘）是特定的。

是。Hifi_reads.bam 格式继续遵守 SAM 规格。为提高数据量效率，Revio BAM 格式在碱基质量编码与 read 排序次序方面存在细微差异。

是。示例数据集位于此处。

  1. 显示数据针对用于 HG002/GM24385 的单 Revio SMRT Cell 芯片。
  2. 基于 HG002/GM24385 人类 DNA 的 HiFi 产出规格使用 Nanobind CBB 试剂盒提取，并用 SMRTbell 制备试剂盒 3.0 制备。
  3. 运行时间规格针对测序反应而言。
  4. 年通量基于 1,300 块 Revio SMRT Cell 芯片计算。
  5. 期望覆盖率与通量均为估计值。因文库质量与片段长度差异，覆盖率可能会有所不同。
  6. HiFi产量取决于于文库片段的大小。对于小于15 kb的文库，产量通常较低。

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