1st July 2024

sparQ DNA Library Prep Kit – Prepare for next generation sequencing success with the highest quality library.

Features & Benefits

Simplified 2-step protocol speeds sample prep to under 3 hours and minimizes sample loss from transfer steps

  • Increased library yields enable the construction of low input DNA sample from 250 pg
  • Minimized bias improves coverage across difficult to sequence regions ensuring optimal results and reduced coverage gaps\
  • PCR-free workflows enabled from 100 ng input DNA
  • Improved overall sequencing workflow economics

sparQ DNA Library Prep Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

sparQ DNA Library Prep Kit is optimized for the rapid construction of DNA libraries from fragmented double-stranded DNA for sequencing on Illumina® NGS platforms. DNA polishing reactions are combined in a single step to convert fragmented DNA into 5′-phosphorylated and 3′-dA-tailed DNA fragments suitable for direct ligation of sequencing adapters without the need for an intervening cleanup saving valuable time and bead purification expense. The HiFi PCR Master Mix and Primer Mix allows for the optional, unbiased amplification of fragments with appropriate adapters ligated to both ends. The kit is compatible with input amounts from 250 pg to 1 μg DNA and multiple sample types.

Streamlined workflow

  • Two step protocol generates sequencing ready library in 2.5 hours
  • Fewer transfer steps minimizes sample loss

The streamlined workflow can be completed in under 3 hours with minimal hands-on time. A single tube is used for DNA polishing, ligation, and cleanup. A second tube is used for workflows requiring PCR amplification and a final tube receives the sequencing-ready library.

56% Increase in library yields

  • Highest library yields from enzymes engineered for optimal sensitivity and efficiency
  • 250 pg to 1 µg of input DNA produces unbiased libraries
  • PCR-Free workflows starting with 100 ng input DNA

The streamlined workflow can be completed in under 3 hours with minimal hands-on time. A single tube is used for DNA polishing, ligation, and cleanup. A second tube is used for workflows requiring PCR amplification and a final tube receives the sequencing-ready library.

Minimize bias and gaps in your sample genome coverage

  • High fidelity master mix is formulated to increase yield even in GC and AT-rich challenging regions
  • HiFi efficiency reduces the number of amplification cycles required to create sequence ready libraries
  • Potential for PCR-derived artifacts is significantly reduced

Amplified libraries were prepared from 100 ng of microbial genomic DNA and subsequently sequenced on Illumina MiSeq. 2 million reads from each tested library were down-sampled and analyzed. Coverage uniformity for different library preparation kits were compared by plotting normalized coverage for both extreme AT-rich regions (8%-20% GC-content) and GC-rich regions (75%-88% GC-content).

Create amplified libraries with PCR-Free results

  • Low amplification bias enables better coverage uniformity
  • Coverage of amplified workflow is matched to coverage of PCR-Free workflow

Library amplification with sparQ HiFi PCR Master Mix contained in the sparQ DNA Library Prep Kit resulted in uniform coverage across the wide range of GC-content. Libraries were prepared by using sparQ DNA Library Prep Kit with 100 ng input DNA. Coverage depth against GC-content of libraries amplified by sparQ HiFi PCR Master Mix (orange) were compared to corresponding libraries without amplification (dark blue: PCR-free library). GC content distribution of targeted genomes is indicated by grey line.

Improve your sequencing results and economics

  • Obtain a comprehensive view of your sample genome
  • Industry leading sequencing results
  • High mapped reads with minimal duplication rates


sparQ DNA Library Prep Kit generates high quality DNA libraries with minimal duplication rates. Libraries were prepared with 1 ng and 100 ng of microbial genomic DNA and subsequently sequenced on Illumina MiSeq. Each library was down-sampled to 2 million reads (150 bp paired-end reads) and aligned to a reference genome with only unique alignments reported.

sparQ Adapter Barcode Sets

sparQ Adapter Barcode Sets A and B are single index barcoded oligonucleotide adapters designed for use with the Quantabio sparQ DNA Library Prep Kits in the construction of libraries for Illumina NGS platforms. Each sparQ Adapter Barcode Set contains 12 single index barcodes for 96 rxns.

Contents:

sparQ DNA Library Prep Kit contains:

  • DNA Polishing Enzyme Mix
  • DNA Polishing Buffer
  • DNA Ligase
  • DNA Rapid Ligation Buffer
  • HiFi PCR Master Mix (2X)
  • Primer Mix
  • sparQ Adapter Barcode Sets contain twelve adapters with distinct barcode sequences (*see above)

Storage & Handling:

Store kit components in a constant temperature freezer at -25°C to -15°C upon receipt. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.

Resources:

Ordering:

  • sparQ DNA Library Prep Kit 24 rxns: 95191-024
  • sparQ DNA Library Prep Kit 96 rxns: 95191-096
  • sparQ Adapter Barcode Set A, 12 single index barcodes for 96 rxns: 95193-A96
  • sparQ Adapter Barcode Set B, 12 single index barcodes for 96 rxns: 95193-B96