Onso was the second sequencer Pacific Biosciences (PacBio) revealed during this year’s American Society of Human Genetics meeting (read about the first sequencer here). This benchtop sequencer uses sequencing by binding (SBB), a method unlike any other instrument on the market. SBB incorporates native nucleotides, has reduced molecular scarring, and reportedly results in significantly higher accuracy base calls than traditional short-read sequencers. The following sections highlight the important specifications of this unique device.
Read type and length
Onso is PacBio’s first and only short-read sequencer. This new instrument has two available reagent kits (200- and 300-cycle). The read lengths can be configured at 2 x 100 bp or 1 x 200 bp with a 200-cycle kit, or 2 x 150 bp with a 300-cycle kit. Both options have high quality scores of ≥90% Q40+.
Flow cell/SMRT Cell options
Instead of a SMRT Cell, like other PacBio sequencers, Onso operates on a single, dual-lane flow cell. Prior to initiating the sequencing run, each lane can be loaded with separate pools of single or multiplexed samples.
Data output
The data output is 80–100 Gb using a 200-cycle kit with an expected run time of 32 hours, while the 300-cycle kit has a data output of 120–150 Gb with a 48-hour run time. The number of expected reads for each sequencing run is 400–500 M.
Analysis options
FASTQ generation can be completed on the instrument and demultiplexing is also expected to be offered onboard during the final release. The resulting data can then be easily combined with many existing tools for secondary analysis.
Common applications
• Cancer screening (cfDNA) and research
• Single-cell sequencing
• Whole exome sequencing
• Biomarker discovery
Release date
Those interested in Onso can place orders with PacBio and shipments are anticipated for the first half of 2023.
Highlights
• The first short-read sequencer from PacBio and a compact benchtop instrument.
• Uses sequencing by binding, a unique short-read method with an increased sensitivity that lowers the limit of detection.
• Collaborations with 10x Genomics, Twist Biosciences, and other well-known vendors to streamline protocols onto the new device.
• Ultra-high Q40+ data quality allows for a decrease in read depth needed for accurate analysis.
• Low duplication rates, minimal index-hopping, and the ability to fully resolve low-complexity regions better than SBS technology.
To compare sequencing technologies please visit Biocompare
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About the Author
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Benjamin Atha holds a B.A. in biology from Hood College and an M.S. in biological sciences from Towson University. With over 9 years of hands-on laboratory experience, he's well-versed in next-generation sequencing systems. Ben is currently the editor for SEQanswers.
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