PacBio SMRTbell® 3.0 Library Prep, automated on Callisto™
The PacBio SMRTbell 3.0 Library Prep App converts DNA into SMRTbell libraries at a miniaturized reaction scale on Callisto, for HiFi long-read sequencing.
Callisto™ is For Research Use Only. Not for use in diagnostic procedures. Oxford Nanopore products are RUO. Regulatory status and claims regarding third-party products are the responsibility of the third party.
How it runs on Callisto
Load
Load DNA with the PacBio SMRTbell prep kit 3.0 reagents.
End repair and ligation
Callisto runs end repair and SMRTbell adapter ligation walk-away.
Nuclease and size selection
Nuclease treatment and optional bead-based size selection run on deck.
HiFi libraries out
SMRTbell libraries ready for Revio or Sequel IIe.
Run a new app on the Callisto™ you already have.
The protocol is already validated and automated.
Load the kit and walk away.
Performance data
Miniaturized PacBio preps without compromising on performance
Miniaturized half-scale PacBio libraries prepared on Callisto yield equivalent final library masses to full-scale manual preps. Library yield scales reliably with input gDNA (1–5 µg), demonstrating robust and reproducible performance across a broad input range.
Robust size selection to eliminate short fragments
The PacBio SPK 3.0 library prep App on Callisto is optimized for robust size selection performance to minimize the number of short fragments (<5 kb), which is critical for maximizing HiFi yields on PacBio sequencers. This minimizes the need for other size selection procedures for the input DNA (such as Short Read Elimination or SRE) and prepared libraries (such as BluePippin). Performing Short Read Elimination (SRE) on the DNA input for the App delivers only modest gains in eliminating short fragments over un-SREd DNA, suggesting excellent size selection performance by the App.
Consistent, high-quality HiFi sequencing on Revio
Callisto-prepped libraries deliver high-quality HiFi sequencing results on PacBio Revio, performing on par with or better than manual preparations. Key sequencing metrics—including HiFi read N50, total read count, and Q score—are maintained with high consistency. Data presented includes 12 manually prepared libraries and 24 libraries processed using Callisto.
Robust variant calling performance
Callisto-prepped libraries deliver highly accurate variant calling results, matching or exceeding manual preps in SNP and INDEL precision, recall, and F1 scores. Libraries were prepared using HG001 reference DNA and sequenced on PacBio Revio. Variant calling was performed with Google DeepVariant and benchmarked against hg38 NIST v4.2.1 using hap.py.
Source: results previously published on this page. App note: PacBio SMRTbell 3.0 Library Prep.
Frequently asked questions
What DNA input does the PacBio SMRTbell 3.0 App need?
1 to 5 µg of DNA.
Which steps does Callisto automate?
End repair, ligation, nuclease treatment and optional size selection.
How many samples can I prep per run?
Flexible batch sizes from 4 to 24 reactions in a single run.
What applications is the app used for?
De novo genome assembly, variant detection, AAV sequencing and amplicon sequencing.
See PacBio SMRTbell 3.0 DNA Library Prep on a Callisto in your lab.
Book a demo, or browse the full Callisto app menu.
Callisto™ is For Research Use Only. Not for use in diagnostic procedures. Oxford Nanopore products are RUO. Regulatory status and claims regarding third-party products are the responsibility of the third party.