## Maintain rapid turnaround time through flexible batch sizing

Volta's PacBio™ SMRTbell 3.0 Library Prep App offers an automated process of converting DNA into SMRTbell libraries, at a miniature reaction scale. The process includes end repair, ligation, nuclease treatment and optional size selection. The App can be used for de novo genome assembly, variant detection, AAV sequencing and amplicon sequencing.

### KEY BENEFITS

- Get 5.5 hours of walkaway time for 24 samples
- Prep 2x more reactions from library prep kit
- Achieve the highest quality libraries with no prior experience preparing libraries

## App Specifications

**Input:** 1 - 5 µg DNA  
**Output:** HiFi libraries for Revio/Sequell IIe systems  
**Hands-on time:** Less than 30 mins  
**Walk-away time:** 5.5 hours  
**Throughput:** Flexible, 4 to 24 rxns in a single run  
**Chemistry:** PacBio SMRTbell prep kit 3.0 (102-182-700) with bead-based size selection

## Miniaturized PacBio preps without compromising on performance

Miniaturized half-scale PacBio libraries prepared on Callisto (blue) yield equivalent final library masses to full-scale manual preps (grey). 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**

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, green trace) on the DNA input for the App delivers only modest gains in eliminating short fragments over un-SREd DNA (blue trace), 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, ensuring reliable and high-fidelity data with every run. Data presented includes 12 manually prepared libraries and 24 libraries processed using Callisto.

## Robust variant calling performance

Callisto-prepped libraries (blue) deliver highly accurate variant calling results, matching or exceeding manual preps (grey) 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.

To process samples using PacBio SMRTbell 3.0 Library Prep you will need the following materials:

- Volta Labs
- DNA Library Prep for PacBio® (SMRTbell® 3.0) - 48 rxns Accessory Kit
- LP0001 PacBio*
- PacBio SMRTbell prep kit 3.0
- 102-141-700
- PacBio AMPure® PB beads
- 100-265-900
- PacBio PacBio Elution Buffer#
- 101-633-500
- PacBio SMRTbell cleanup beads#
- 102-158-300
- Low TE Buffer pH 8.0#
- 102-178-400

*Please contact our partners directly to purchase these reagents.

#The SMRTbell prep kit 3.0 does not include SMRTbell cleanup beads, AMPure® PB beads, low TE Buffer and a sufficient quantity of PacBio Elution Buffer to perform sample preparation. Purchasing these reagents separately from PacBio is required.

To order: sales@voltalabs.co

## PacBio Compatible Partner

Volta Labs is partnering with PacBio to offer walk-away automation for PacBio library prep, enabling labs to scale sample prep for sequencing. Through preloaded apps on the Callisto Sample Prep System, PacBio customers are able to prepare high-quality PacBio libraries with less than 30 minutes of hands-on time and 5.5 hours of walk-away time. Users can be trained in minutes to generate PacBio libraries with minimal inter-operator variability.