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

Streamlined Workflow

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

56% Increase in Library Yields

Library Yield Analysis

    • 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

Minimize bias and gaps in your sample genome coverage

Coverage Across Challenging Regions

    • 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

Create amplified libraries with PCR-Free results

Create amplified libraries with PCR-free resultsRegions

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

Improve your sequencing results and economics

Improve Sequencing Results and Economics

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

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.

sparQ Adapter Barcode Set A           sparQ Adapter Barcode Set B
Barcode Number    Barcode Sequence Barcode Number  Barcode Sequence
2 CGATGT         1 ATCACG
4 TGACCA   3 TTAGGC
5 ACAGTG   8 ACTTGA
6 GCCAAT   9 GATCAG
7 CAGATC   10 TAGCTT
12 CTTGTA   11 GGCTAC
13 AGTCAA   20 GTGGCC
14 AGTTCC   21 GTTTCG
15 ATGTCA   22 CGTACG
16 CCGTCC   23 GAGTGG
18 GTCCGC   25 ACTGAT
19 GTGAAA   27 ATTCCT

 

Performance Data

sparQ DNA Library Prep Workflow


sparQ DNA Library Prep Workflow

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.


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Coverage Across Wide Range of GC-Content


Uniform coverage across wide range of GC-content

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 gray line.


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Improve Sequencing Results and Economics


Improve Sequencing Results and Economics

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.


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Coverage Across Challenging Regions


Coverage Across Challenging Regions

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).


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sparQ DNA Library Prep Kit Yield Analysis


sparQ DNA Library Prep Kit Yield Analysis

sparQ DNA Library Prep Kit produces high quality libraries from a broad range of DNA inputs with significantly higher yields. Libraries were prepared with Covaris-sheared DNA (250 bp average size) using kit manufacturers’ instructions. Amplified libraries (6 amplification cycles for 100 ng input DNA and 13 amplification cycles for 1 ng input DNA) were quantified with Qubit fluorometric quantitation method.


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Details

  • 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 are in each set.

    sparQ Adapter Barcode Set A            sparQ Adapter Barcode Set B
    Barcode Number    Barcode Sequence Barcode Number  Barcode Sequence
    2 CGATGT         1 ATCACG
    4 TGACCA   3 TTAGGC
    5 ACAGTG   8 ACTTGA
    6 GCCAAT   9 GATCAG
    7 CAGATC   10 TAGCTT
    12 CTTGTA   11 GGCTAC
    13 AGTCAA   20 GTGGCC
    14 AGTTCC   21 GTTTCG
    15 ATGTCA   22 CGTACG
    16 CCGTCC   23 GAGTGG
    18 GTCCGC   25 ACTGAT
    19 GTGAAA   27 ATTCCT
  • Storage & Handling
    Store components in a constant temperature freezer at -25°C to -15°C upon receipt. Refer to the product label for applicable expiration date.
  • Related Resources