LongPlex™ Long Fragment Multiplexing Kit with Index Set 1

Realize the potential of your PacBio long read sequencer

Product Description

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.

• Plate-based, enzymatic method to simultaneously fragment & index genomic DNA for scalable PacBio HiFi sequencing
• Massive multiplexing using 96 unique dual indexes (UDI) that can be combinatorially expanded with PacBio indexing
• Early sample pooling greatly reduces all-in cost per sample for long read sequencing without sacrificing data quality
• Run more long read samples per flow cell without compromising PacBio HiFi sequencing quality
• Supports long read applications including microbial and small genome sequencing, metagenomics, low pass sequencing, and targeted hybrid capture
  
  

Transposase-based gDNA fragmentation ensures a consistent, scalable generation of 8 -10 kB fragment lengths. The 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.

Figure 1: 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. 

Kit Content

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

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

LongPlex XL Workflow

A modified LongPlex™ Protocol (LongPlex XL) for generation of HiFi Read Lengths >10 kb using PacBio Short Read Eliminator (SRE) is available. Find more details about this improved long-read tagmentation workflow on the LongPlex XL poster and contact us to receive the protocol.

Reducing Long Read Sequencing Costs Through Massive Multiplexing

Efficiently multiplex more samples per flow cell without compromising sequencing quality on PacBio HiFi long-read sequencers.  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.

Specifications

*Other sample types may be compatible. Please contact BioCat 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

Additional Resources

Scalable Long-Read Low-Coverage Human Whole Genome Sequencing (lcWGS) for Genotype Imputation (poster)

An Efficient Enzyme-Based Library Workflow for Multiplexed Long Read Whole Genome Sequencing (poster)