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LongPlex Launch Celebration

Let’s celebrate making long-read sequencing scalability a reality.

October 8, 2024

8:30 am PDT   l   11:30 am EDT   l   16:30 CET

Join this virtual event to see the LongPlex workflow in action.  Hear from 3 early users about how they applied LongPlex for microbial WGS and low pass long read sequencing.

Agenda & Registration

Key Benefits: Realize the potential of your PacBio long read sequencer

Realize the throughput and cost-effectiveness of PacBio HiFi sequencing using seqWell’s LongPlex Long Fragment Multiplexing Kit.  Its speed, simplicity, and scalability enable massive sample multiplexing that unleashes scalable long read sequencing.

  • SPEED: Eliminate mechanical shearing using a fully scalable, enzymatic method to simultaneously fragment & tag genomic DNA
  • SIMPLICITY: Automation-friendly, plate-based method with total workflow performed in <2 hours (30 minutes hands-on time)
  • SCALABILITY: Massive multiplexing using 96 unique dual indexes (UDI) that can be combinatorially expanded with PacBio indexing
  • SAVINGS: Early sample pooling greatly reduces all-in cost per sample for long read sequencing without sacrificing data quality

Transposase-based gDNA Fragmentation

Consistent, scalable generation of 8 – 10 kB fragment lengths

Enzymatic fragmentation using adapter-loaded transposase allows for simultaneous fragmentation and indexing of gDNA.  Importantly, this eliminates the need for mechanical shearing of gDNA and greatly increases sample throughput.

DNA samples were analyzed on the Femto Pulse system following LongPlex processing using the standard PCR-free protocol and SMRTbell 3.0 library preparation.  The traces show reproducible fragmentation profiles across four 24-plexes (full plate of 96 samples pooled in 4 separate pools of 24) with an average fragment length >8 kb. 

Simple Workflow for Scalable Fragmentation and Multiplexing

 

  • Simultaneously fragment and index up to 96 gDNA samples with no DNA shearing required
  • Plate-based protocol performed in < 2 hours with only 30 minutes of hands-on time
  • Produces fragments ranging from 8 – 10 kb long
  • Sample multiplexing via pooling of up to 96 samples prior to SMRTbell library preparation
  • PCR-free and PCR-based protocols available to support a variety of needs including difficult samples

 

Reducing Long Read Sequencing Costs Through Massive Multiplexing

Efficiently multiplex more samples per flow cell without compromising sequencing quality on PacBio sequencing platforms.  To run 384 long read samples, you no longer need to perform 384 PacBio SMRTbell library preparations! Simply use the plate-based LongPlex method, followed by 4 SMRTbell library preps enabling 384 sample on a single flow cell.  Pooling samples early allows you to maximize sequencer throughput while driving down costs.

 

LongPlex Specifications

Specs Description
Primary Applications Long read sequencing for:

  • Microbial & small whole genome sequencing
  • Low pass sequencing
  • Metagenomics
  • Targeted hybrid capture
Sample Types* Genomic DNA (DIN ≥8.0 recommended)
Reactions per Kit 96 reactions
DNA Input Recommended 250 – 500ng
Indexing Method Combinatorial Dual Indexing
Number of Unique Index Combinations 96
Batch Size** 1-24 samples
Output Fragment Range*** 5,000 – 8,500bp
PCR Amplification
  • WGS: PCR free & PCR-plus methods available
  • Hybrid Capture: 8-10 PCR cycles recommended
Total Protocol Time
  • PCR-free: 60 minutes (20 minutes hands-on)
  • PCR-plus: 3-4 hours including long range PCR (30 minutes hands-on)
Sequencer Compatibility PacBio Revio

PacBio Sequel II & IIe

*Other sample types may be compatible. Contact seqWell’s support for guidance. **Up to 96 samples can be pooled past LongPlex barcoding prior to PacBio SMRTbell but it may impact read output and coverage uniformity ***Fragment size will depend on magnetic bead cleanup ratios used

LongPlex Long Fragment Multiplexing Kit Includes:

  • Tagging Reagent Plate (96-well microtiter plate)
  • 3X Coding Buffer
  • X Solution
  • Library Primary Mix for optional PCR
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