Technical note

SIZE SELECTION OF PACBIO SMRTBELL LIBRARIES WITH
THE LIGHTBENCH INSTRUMENT FROM YOURGENE HEALTH
Introduction
Precise size selection is essential for maximizing the benefitsof PacBio® highly accurate long-read sequencing. TheLightBench® from Yourgene Health uses Ranger® Technology todeliver accurate size selection for whole genome sequencing
and includes the ability to perform fragment length analysis
for additional quality control. With an automated workflow tooptimize walk-away time, the LightBench® offers a scalablesize selection solution for the preparation of SMRTbell® gDNA
libraries for HiFi sequencing.

In experimental benchmarking of 84 human gDNA samples
from fresh blood for library preparation for HiFi long-readsequencing, the LightBench® achieved consistent yields for
8 kb or 10 kb cutoffs across all samples and a 0% failure rate.
The 8 kb cutoff yielded an average recovery of ~83% of the
input SMRTbell library while the 10 kb cutoff recovered anaverage of ~67%.

AB

Input DNA

500 ng–2.0 µg

Throughput

12 samples

Run time~3 hours

Hands-on time~20 minutes

8 kb cutoff~83% recovery *10 kb cutoff~67% recovery

Pre-size selection

Post-size selection**Size selection recovery of SMRTbell library.

This technical note provides an overview of the size selection
workflow for PacBio SMRTbell sequencing libraries with theLightBench® and reports experimental results demonstrating
effective size selection at two fragment lengths.

Figure 1. Representative HiFi read length distribution (A) and comparison of pre- andpost-size selection on the Agilent Femto Pulse system (B) for 10 kb cutoff demonstrates
effective removal of <10 kb fragments.

Workflow overview
Required materials and equipment38238616000

18000

Part numberLightBench®

CG-12500-03

Materials

614000Equipment

20000

22000

Cassette and Dual Dye
Loading Buffer kit

32400026000

Figure 2. Mean read length distribution across 84 samples on the Revio™ system.

Following library preparation and size selection with theLightBench®, 84 samples were sequenced on the Revio™
platform. Mean HiFi read length was 18,517.5 bp (figure 2,max: 25,613 bp; min: 14,202 bp) and mean HiFi data yield
was 101.27 Gb/SMRT® Cell (figure 3).

CG-14100-13-050-31-31

IncludesDual Dye Loading Buffer
7 kb + 7 kb markers

CG-14000-31-31

0.5% In-Channel Filter
Size-selection cassette

CG-10600-13-050

Workflow steps
1. Prepare the Ranger® Technology software setup according
to Yourgene LightBench® recommendations.
2. Prepare the samples by combining with Dual Dye Loading
Buffer containing the 7 kb marker (CG-14000-31-31). Mix
thoroughly to homogenize and spin down to remove any
air bubbles.
3. Prepare the cassette and remove excess buffer from thereservoir.

3119814011260806100

4. To load samples into the cassettes, remove recommended
volume of buffer from each loading well, and dispense thefull volume of aspirated samples into loading wells.

1783120

5. Allow the electrophoresis process to proceed.

1140

Figure 3. HiFi yield (Gb) distribution across 84 libraries on the Revio system.

These results demonstrate the LightBench® as an effective
method for the size selection of HiFi libraries. See theworkflow overview for a summary of the recommended size
selection procedures and visit Yourgene Health for detailed
protocol steps.

6. For sample extraction, remove the cassette from theinstrument and rinse the extraction wells.
7. Refill extraction wells with 1×TBE buffer and insert theIn-Channel Filter (ICF) array into the extraction well. Fill ICF
collection chambers with 1×TBE buffer.
8. Return the cassette to the LightBench® drawer tray andallow the extraction process to proceed. Extract targetedmaterial when prompted by the software.
9. Size selected fractions should be bead-cleaned andassessed for suitable concentration and fragment length
distribution prior to sequencing.

To learn more, visit yourgenehealth.com
This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by Yourgene Health. For more information about commercial rights, please email info@yourgenehealth.com.
Research use only. Not for use in diagnostic procedures. © 2023 Pacific Biosciences of California, Inc. (“PacBio”). All rights reserved. Information in this document is subject to change without notice. PacBio
assumes no responsibility for any errors or omissions in this document. Certain notices, terms, conditions and/or use restrictions may pertain to your use of PacBio products and/or third-party products. Refer tothe applicable PacBio terms and conditions of sale and to the applicable license terms at pacb.com/license. Pacific Biosciences, the PacBio logo, PacBio, Circulomics, Omniome, SMRT, SMRTbell, Iso-Seq, Sequel,Nanobind, SBB, Revio, Onso, Apton, and Kinnex are trademarks of PacBio. Ranger® Technology and LightBench® are registered trademarks of Yourgene Health.

102-326-596 REV01 16NOV2023