BioXp™ System Next Generation Sequencing Library Construction Kits

The BioXp™ 3200 System automates the construction of next generation sequencing libraries. This genomic workstation provides a simple process that eliminates the time, hassle, and potential for variability that can occur with manually prepared NGS libraries. Built for Illumina® sequencing platforms, the BioXp™ 3200 NGS library construction method allows the construction of up to 16 barcoded libraries starting with 10 ng to 50 ng of input genomic or plasmid DNA. This kit is also available in a non-automated format.

The BioXp™ System NGS Library Construction Kits offer:

  •  Simple, hands-free NGS library construction
  •  Complete kit includes barcodes and magnetic beads
  •  Fast creation of 8 or 16 barcoded libraries in under 5 hours
  •  Ideal for sequencing of genomic and plasmid DNA
  •  An optimized protocol for flexible input DNA (10-50 ng)
  •  Ready to use kits in automated and non-automated formats
The BioXp™ 3200 system is a compact genomic workstation that enables a variety of hands-free molecular biology applications.

Built for ease of use, even novice scientists can successfully prepare a library ready for Illumina NGS sequencing. Simply add sheared DNA sample to the sample plate, load the reagents on the BioXp™ deck and press start.

The BioXp™ system runs hands-free to create barcoded NGS libraries ready for normalization, pooling, and sequencing in under 5 hours. The reliable NGS library construction chemistry efficiently adds to adapters to input DNA samples from 29% to 68% GC content to produce large numbers of unique sequencing reads.

Using the BioXp™ system and the NGS Library Construction Kits, you can consistently build reliable NGS libraries. This easy-to-use system is ideal for labs needing automated low to medium throughput of NGS library preparation. The BioXp™ system is a complete genomic workstation that enables other molecular biology applications including generation of synthetic double stranded DNA fragments and automated cloning. Learn more about the BioXp™ 3200 System.

Non-Automated NGS Library Construction Kit

Not ready for automation? Want to compare this library construction method to your present method? The BioXp™ NGS Library Construction Kit is also available as a manual library construction format. In either case our optimized enzymes and buffers facilitate the ligation of the provided barcode adapters resulting in diverse sequencing reads. In either an automated or manual format, these complete NGS library construction kits include barcode adapters and magnetic beads so you have all the key components to ensure the highest quality libraries for reliable sequencing results.

Other Available BioXp™ products:

BioXp™ Tiles

BioXp™ pUCGA 1.0 Cloning

BioXp™ Custom Cloning

How It Works

All key reagents required to perform automated NGS library construction on the BioXp™ system are provided in the kit.

  • Shear the DNA using your method of choice
  • Add the sheared DNA sample to the sample plate
  • Place the BioXp™ reagents, fresh 70% ethanol and tips onto the BioXp™ deck
  • Close the BioXp™ system and press start
  • Collect material when complete
  • Normalize, Pool and Sequence your Library
BioXp™ 3200 NGS Library Construction Kit workflow. Starting with sheared genomic DNA, the BioXp™ 3200 System allows automation of the NGS library construction process resulting in bar-coded libraries ready for normalization, pooling and sequencing. This same chemistry is available in a non-automated format for those without access to a BioXp™ system.

Simple and rapid set up of BioXp™ 3200 for NGS library construction. The BioXp™ 3200 System deck, with all NGS library construction reagents in appropriate location is shown. Setup for automated library construction can be accomplished by loading kit components as described here. First, add the sheared DNA sample to the sample plate and place in the location shown. Next place the pre-aliquoted NGS library construction reagents, the barcode plate, and magnetic bead strips provided with the kit, in the locations shown. Fill the ethanol reservoir and place in the location shown. Finally, add the transfer tips into the holders (tips sold separately), close the lid and press start

Performance

Robust ligation of adapters

The robust addition of adapter sequences to sheared input genomic DNA. A sample of sheared input Vibrio natriegens genomic DNA and the final prepared library were run an Agilent 2100 Bioanalyzer and a High Sensitivity DNA Chip. The resulting Bioanalyzer gel image and electropherograms are shown. Input and final prepared library are indicated by red and blue lines, respectively, in the electropherogram. A red arrow indicates average size of 247 bp for input DNA and the blue arrow indicates expected size shift of peak of 333 bp in the final prepared library.

Reliable chemistry consistently delivery low % of adapter dimers and large numbers of mapped reads

Construct Results

Sample Average Fragment Size % Dimer % Library # Reads % Mapped Average Sequencing Coverage Max Cvg
Bacillus-1 342 2% 92% 1,700,924 96% 43.92 250
Bacillus-2 322 1% 92% 1,623,244 96% 42.34 199
Bacillus-3 324 5% 90% 1,375,190 88% 35.94 489
Bacillus-4 339 1% 92% 1,793,744 98% 48.87 343
Vibrio-1 341 0% 87% 1,743,414 98% 39.51 102
Vibrio-2 328 2% 91% 1,664,782 95% 36.38 105
Vibrio-3 333 3% 90% 1,772,470 94% 36.79 108
Vibrio-4 333 1% 92% 2,929,992 97% 65.62 158

Sample result from BioXp™3200 NGS library construction and sequencing. Replicate NGS libraries were constructed from samples of Bacillus genomic DNA or Vibrio genomic DNA using the BioXp™ 3200 NGS Library Construction Kit and BioXp™ instrument. The average size of fragment sequenced and % dimer present in the final library and the percent library were determined using an Agilent 2100 Bioanalyzer and a High Sensitivity DNA Chip. Resulting libraries were sequenced using Illumina MiSeq® system. Total number of reads, the percentage of reads mapped to reference genome, average coverage (average number of reads that map to reference genome), and maximum coverage were determined using the CLC Genomics Workbench. Data shown indicate creation of a high-quality library that resulted in reliable sequencing data output.

Reliably assemble libraries from DNA samples from 29% to 68% GC content

Minimal GC bias for reliable results across a range of samples. GC bias for three different bacterial genomes with a range of GC content [Low, C. difficile 29% GC (left), Moderate V. natriegens 45% GC (middle) and High, B. pertussis 68% GC (right)] were used to prepare NGS libraries using the BioXp™ 3200 NGS Library Construction Kit. For each genome, the GC content of the reference genome in 100 bp bins was plotted vs. the normalized coverage across these bins using Picard CollectGCBiasMetrics. In the absence of sequencing bias, all bins would be equally represented and indicated by a horizontal distribution centered on a normalized coverage of 1. In the figure shown, distribution of GC content in the genome is indicated by the blue histograms. The normalized coverage for the BioXp™ NGS Library Construction Kit is shown by the orange line and is consistent with the level of bias reported for other commercially available kits.

BioXp Automated and Manual NGS Library Construction Kits

Products Catalog # Pack Size List Price
BioXp™ 3200 NGS Library Construction Kit* BX2000-08 8 rxns $399
BioXp™ 3200 NGS Library Construction Kit* BX2000-16 16 rxns $749
NGS Library Construction Kit, non-automated** BX2000-08M 8 rxns $379

*Automated reagent kit. Requires the BioXp™ 3200 System to run chemistry.

**Non-automated version of kit utilizes same enzymes, buffers, magnetic beads and barcodes as automated version


Kit Components

Component BX2000-08 BX2000-16 Storage Contents/Use
NGS Library Construction Reagent Block 1 1 -20° C Contains enzymes, buffers used for NGS library preparation
Barcode Plate 1 1 -20° C Source of barcode adapters
Magnetic Bead Purification Strip 1 2 4° C Magnetic beads for size selection and reaction clean-up
Sample Plate 1 1 4° C or -20° C Used for addition of sheared input DNA and where final constructed library is stored upon run completion

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