sparQ DNA Library Prep Kit

Streamlined, versatile single-tube solution for high quality library rep

Features & Benefits

  • Fast, easy single-tube solution completes library prep in 2.5 hours
  • Suitable for a wide range of input amounts from as low as 250 pg
  • Optimized chemistry ensuring superior library prep sensitivity and efficiency
  • Higher library yields compared to other library prep kits
  • High efficiency enables PCR-free workflow from 100 ng input

 

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. The simplified protocol speeds up library prep to 2.5 hours with minimal hands-on time and accommodates DNA input amounts from 250 pg to 1 μg. DNA polishing reactions are streamlined into a single step to convert fragmented DNA into 5'-phosphorylated and 3'-dA-tailed DNA fragments. This is followed by high efficiency adapter ligation in the same tube. PCR-free workflows are enabled from 100 ng of starting material. If library amplification is required, the HiFi PCR Master Mix and Primer Mix ensure even amplification with minimal bias.

 

Streamlined  workflow

sparQ DNA Library Prep workflow


sparQ DNA Library Prep workflow

The streamlined workflow combines DNA polishing and adapter ligation in a single tube for rapid construction of libraries from fragmented DNA. An optional step using the HiFi PCR Master Mix and Primer Mix ensures even library amplification with minimal bias.


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  • Generates sequencing ready library in 2.5 hours with minimal hands-on time
  • Simplified single-tube solution minimizes sample loss


Higher library yields

Higher library yields


Higher library yields

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 human genomic DNA (250 bp average size) using kit manufacturers’ instructions. Amplified libraries (6 PCR cycles for 100 ng input DNA and 13 PCR cycles for 1 ng input DNA) were quantified with Qubit fluorometric method.


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  • Unique enzymes engineered for optimal sensitivity and efficiency
  • Over 50% increase in library yields than other library prep kits
  • Compatible with a broad range of DNA inputs from 250 pg to 1 μg


Uniform coverage across challenging regions

Coverage across a wide range of GC-content


Coverage across a wide range of GC-content

PCR-free (dark blue) and amplified libraries (orange) were prepared from 100 ng of microbial genomic DNA of different species using sparQ DNA Library Prep Kit. Libraries were sequenced on Illumina MiSeq and 2 million sequencing reads from each library was analyzed. Coverage uniformity was examined by plotting normalized coverages of the library against GC-content of the targeted genome. GC-content distribution of targeted genomes is indicated by gray line. Both PCR-free and amplified libraries resulted in uniform coverage across a wide range of GC-content..


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  • Even coverage across a broad GC-content
  • Coverage of amplified libraries closely resembles coverage of PCR-free workflow


High quality sequencing metrics

Higher quality sequencing metrics


Higher quality sequencing metrics

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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  • High mapping percentage and low duplication artifacts
  • Industry leading sequencing results ensures better return on investments

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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sparQ NGS Library Prep Solutions

Details

  • Contents

     

     

    Volume

    Component Description

    Cap Color

    95191-024

    95191-096

    DNA Polishing Enzyme Mix

    Blue

    1 x 0.24 ml

    1 x 0.96 ml

    DNA Polishing Buffer

    Blue

    1 x 0.12 ml

    1 x 0.48 ml

    DNA Ligase

    Orange

    1 x 0.24 ml

    1 x 0.96 ml

    DNA Rapid Ligation Buffer

    Orange

    1 x 0.48 ml

    2 x 0.96 ml

    HiFi PCR Master Mix (2X)

    White

    1 x 0.60 ml

    2 x 1.25 ml

    Primer Mix

    White

    1 x 0.036 ml

    1 x 0.144 ml

  • 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.
  • Related Resources
    Product Flyers
    Product Manuals
    Technical Notes
    Publications
    Detailed temporal dissection of an enhancer cluster reveals two distinct roles for individual elements
    Henry Thomas, bioRxiv - 2020
    Abstract
    Many genes are regulated by multiple enhancers that often simultaneously activate their target gene. Yet, how individual enhancers collaborate to activate transcription is not well understood. Here, we dissect the functions and interdependencies of five enhancer elements that form a previously identified enhancer cluster and activate the Fgf5 locus during exit from naïve murine pluripotency. Four elements are located downstream of the Fgf5 gene and form a super-enhancer. Each of these elements contributes to Fgf5 induction at a distinct time point of differentiation. The fifth element is located in the first intron of the Fgf5 gene and contributes to Fgf5 expression at every time point by amplifying overall Fgf5 expression levels. This amplifier element strongly accumulates paused RNA Polymerase II but does not give rise to a mature Fgf5 mRNA. By transplanting the amplifier to a different genomic position, we demonstrate that it enriches for high levels of paused RNA Polymerase II autonomously. Based on our data, we propose a model for a mechanism by which RNA Polymerase II accumulation at a novel type of enhancer element, the amplifier, contributes to enhancer collaboration.
    Genomic Insights and Ecological Adaptations of Deep-Subsurface and Near Subsurface Thermococcus Isolates
    Lilja Caitlin Strang, Western CEDAR - 2020
    Abstract
    Members of the Archaeal genus Thermococcus are sulfur-dependent hyperthermophiles found in hydrothermal vents throughout the world. Previous analysis of a Thermococcus culture collection containing isolates from the Juan de Fuca Ridge, Gorda Ridge, and South East Pacific Rise using amplified fragment length polymorphism analysis and multilocus sequence typing revealed a distinct clade of Thermococcus isolated from the 1996 megaplume event at Gorda Ridge, indicating that they originated from a deep-subsurface habitat. The aim of this study was to elucidate the functional adaptations that allow for the survival of the Gorda Ridge clade in a deepsubsurface habitat as compared to representative Thermococcus isolates from shallow subsurface environments. This was accomplished through a pangenomic analysis of representative isolates in this clade and others from this culture collection. The Gorda Ridge megaplume group was enriched for genes relating to DNA repair and stabilization including a predicted endonuclease distantly related to Archaeal Holliday junction resolvase, DNA mismatch repair ATPase mutS, CRISPR/Cas elements, and dnaK (hsp70). The group was also enriched for ABC-type branched-chain amino acid (BCAA) transport system, enzymes for the Shikimate pathway for aromatic amino acid synthesis, as well as TupA for tungstate transport. These findings suggest that Thermococcus inhabiting deep-subsurface fluid reservoir require the added ability to prevent and repair damage to their DNA, presumably due to the energy demands of DNA replication. The enrichment in BCAA and tungstate transporters may indicate the use of an amino acid catabolism pathway followed by fermentation catalyzed by the tungstopterin containing enzymes aldehyde ferredoxin oxidoreductase and alcohol dehydrogenase, suggesting a preference for peptides over carbohydrates as an energy source in the deep-subsurface.
    Draft Genome Sequence of Phyllobacterium endophythicum mTS5, Isolated from Lupinus micranthus in Tunisia
    Zoé Waller, Microbiology Resource Announcements - 2020
    Abstract
    We report here the draft genome sequence of Phyllobacterium endophyticum mTS5, isolated from a Lupinus micranthus root nodule. The genome consists of 5,454,168 bp, with a GC content of 57%, and contains 5,676 protein-coding sequences.
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    Safety Data Sheets (SDS)
    CofA (PSF)
  • Testimonials

    sparQ DNA Library Prep Kit

    "Very easy to use."

    Scientist

    University Sherbrooke

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