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ExpressPlex Library Prep Kit: Your ticket to speed, throughput, efficiency

The simplicity of the ExpressPlex workflow makes it uniquely well-suited for manual, automated, and ultra high throughput library prep from plasmids and PCR products.

Recorded Webinar | Octant Bio

The game-changing capabilities enabled by ExpressPlex were highlighted in a webinar collaboration with the synthetic biology team from Octant Bio, a drug discovery company located in Emeryville, CA. Octant’s Henry Chan, PhD (Synthetic Biology Lead) and Bryan Jiang (Research Associate) discussed the application of ExpressPlex to OCTOPUS, the high-throughput plasmid sequencing platform that powers Octant’s therapeutic discovery platform. Watch the video to discover how ExpressPlex has dramatically improved OCTOPUS and accelerated their overall discovery process.

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ExpressPlex is a Game Changer for our Customers

“ExpressPlex is literally faster than Sanger. This changes everything for us. Basically, taking a two-day process to one day dramatically shortens time to data.” – Henry Chan, Ph.D., Synthetic Biology Lead, Octant Bio 

“With the simplified and faster library prep that ExpressPlex allowed, it made my life a lot easier.” – Bryan Jiang, Research Associate, Octant Bio

Benefits of using ExpressPlex

With the significantly shortened hands-on time, you can spend your time on data and results, not pipetting. Other benefits of ExpressPlex include:

  • Go from extracted samples to libraries on the sequencer in < 1/2 a day
  • Sequence more samples for less
  • Easily automate your protocol
  • Train any lab tech, minimize chance for error
  • Multiplex up to 1,536 samples
  • Reduce labor while increasing efficiency
  • Decrease chance for errors via minimal handling steps
  • Save significant time, cost, and plastics over other protocols

ExpressPlex One-Step Workflow

ExpressPlex™ Library Prep Kit Single-Step Workflow Demo for High-Throughput Multiplexed Sample Preparation

In this video, we walk you through the single-step workflow to demonstrate the speed and simplicity of the auto-normalizing ExpressPlex technology. The ExpressPlex Library Prep Kit is an ideal solution for high-throughput multiplexed sample preparation for plasmid and amplicon sequencing.

Product Highlights

Auto-Normalization of Read Depth

ExpressPlex demonstrates significantly higher levels of normalization compared to competitors, enabling a simplified workflow where individual normalization is no longer required to achieve more consistent read-depths across samples.

Libraries were prepared from three different reference plasmids of varying sizes across a 10-fold input range at full reaction volume for ExpressPlex (16 µL) and Nextera XT (50 µL), as well as at ¼ miniaturized reaction volume for Nextera XT.

For each method, inputs ranged from 4-40ng for ExpressPlex (standard input is 16ng) and 0.4 – 4ng for Nextera XT (standard input is 1ng).  Inputs were scaled appropriately for 1/4 reactions.

Auto-Normalization of Insert Size

ExpressPlex generates highly consistent library insert sizes over a broad range of DNA inputs.

Libraries from pUC19 plasmid DNA were simultaneously prepared at 3 different input amounts (n=8 each input amount). ExpressPlex demonstrated greater consistency in average insert size across a 10-fold range. In each case, the standard input for both kits (16ng for ExpressPlex, 1ng for Nextera XT) was also included as a mid-range data point.

Success Rates of Plasmid Assembly

Libraries were prepared as described above, sequenced (MiSeq Nano; 2 x150) and de novo assembled using seqWell’s SNAP plasmid assembly pipeline. The sequencing run was downsampled to either 1,000 or 500 average read pairs per sample to identify error-free, circular plasmid assemblies. Even at only 500 average read pairs all ExpressPlex samples generated correct circularized assemblies. At the same number of reads, however, Nextera XT (full and ¼ reactions) failed 8-10% of assembly attempts.

Given the robust read balance and lower number of reads required to generate correct assemblies, ExpressPlex users can multiplex higher numbers of samples on a single Illumina run or utilize a lower capacity flow cell, thereby reducing the cost of sequencing per plasmid.

Confident Amplicon Sequencing

ExpressPlex’s workflow and 10bp barcodes typically provide >99% on-target alignment rates.

20 uniquely mapping amplicons were generated from lambda DNA. The amplicons were used combinatorial as input into the ExpressPlex library prep such that each sample is comprised of 2 lambda amplicons and each well contains a unique amplicon combination. Sample input was maintained at 16 ng and 15 cycles of PCR were used during the library preparation. Following sequencing, demultiplexed reads were mapped to the lambda genome and marked as on-target (expected amplicon combination) or off-target for each well. All wells have an on-target rate >99% (above figure) with an average on-target rate of 99.XX%. Amplicon sequencing with EP enables efficient preparation of up to 1536 amplicons multiplexed on a single lane, enabling high-throughput applications without fear of index-hopping, or PCR chimeras.

Help Save the Environment with Less Plastics

Thanks to the highly efficient ExpressPlex workflow, almost 80% fewer pipette tips and plastics are used to prepare your libraries.

Calculations are based on 96 samples prepared, comparing ExpressPlex to Nextera XT.

ExpressPlex Specifications

Specs Description
Primary Applications* Plasmid and amplicon sequencing
Sample Types* Plasmids, amplicons > 350bp
Reactions per Kit 96 reactions

384 (4 different sets available)

DNA Input Recommended 8 – 40ng
Total Library Prep Time (hands-on time) 90 min

(30 min hands-on)

Indexing Method Combinatorial Dual Indexing
Number of Unique Index Combinations 1,536
Supported Paired Reads (Clusters) per Sample < 4 million (validated to date)
Output Fragment Range** 400 – 1,200bp
Number of PCR Cycles 12 cycles for plasmids
15 cycles for amplicons
Sequencer Compatibility All Illumina sequencing platforms;
Use with Element Biosciences AVITI™ or other sequencing platforms is possible with conversion kits for Illumina libraries

*Other sample types are compatible. Contact seqWell’s application group for guidance.

**Fragment size will depend on magnetic bead cleanup ratios used.

ExpressPlex Library Prep Kit Includes:

  • Indexing Reaction Plate(s)
  • Ready Reaction Mix Plate(s)
  • MAGwise Paramagnetic Beads
  • Users do not need to supply polymerase, primers, or barcodes. These are already pre-mixed into the reagents supplied.