sparQ Universal Library Quant Kit

Fastest qPCR-based library quantification in 40 minutes

Features & Benefits

  • Faster time to results – 50% shorter run time than traditional cycling protocols
  • Accurate, reliable quantification of NGS libraries of various sizes and GC-content
  • High amplification efficiency across a wide linear dynamic range
  • Stabilized, ready-to-use sparQ Universal Fast Mastermix to reduce pipetting steps
  • Superior run to run uniformity ensuring highly precise measurements

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

Description

sparQ Universal Library Quant Kit provides rapid and accurate quantification of libraries prepared for sequencing on Illumina® NGS platforms. Accurate quantification of the number of amplifiable library molecules prior to loading onto a flow cell is a critical step in the NGS workflow and it ensures optimal cluster generation and cost-effective use of sequencing capacity. The sparQ Universal Library Quant Kit uses real-time quantitative PCR (qPCR) to specifically quantify the number of library molecules that possess the appropriate adapter tag at each end.

Accurate library quantification in 40 minutes

Faster run time


Faster run time

Comparison of average qPCR run time for library quantification. sparQ Universal Library Quant Kit uses fast cycling protocol, allowing results to be achieved in 40 minutes versus 1 hour and 20 minutes with the traditional NGS Library Quant Kit.


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  • Optimized for use on any qPCR instrument
  • Reduces qPCR run time by 50%


Complete library quantification solution with unmatched convenience

sparQ Universal Library Quant Workflow


Complete library quantification solution with unmatched convenience

Reactions are prepared by simply adding standard or diluted library sample. Optimized protocols with fast cycling condition are provided for both 10 μl or 20 μl reaction volumes. Following qPCR run, a standard curve is constructed which is then used to determine the concentrations of each unknown diluted library sample.


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  • Supplied with all necessary, ready-to-use reagents to minimize
    pipetting steps
  • Fast cycling protocols work for both 10 μl or 20 μl reaction volumes
  • Reaction setup and data analysis templates and guides provided


qPCR as the most accurate method for library quantification

Comparison of library quantification methods


Comparison of library quantification methods

qPCR is the most sensitive and precise method for library quantification as it measures only DNA fragments containing the appropriate adapter sequences on both ends. Other methods based on spectrophotometry, fluorometry, or microfluidic electrophoresis are prone to variabilities and inaccuracies due to factors such as sensitivity to contaminants or measurement of unsequenceable fragments.


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  • qPCR only quantifies library fragments with adapters attached at both ends
  • Other methods can lead to overestimation of library concentration


Equivalent performance across cyclers: sparQ vs Kapa Library Quantification Kit

Equivalent performance across cyclers: sparQ vs Kapa Library Quantification Kit


Equivalent performance across cyclers: sparQ vs Kapa Library Quantification Kit

Seven Libraries (2 DNA, 5 RNA) were prepared and each quantified using both sparQ Universal Library Quant Kit and Kapa™ Library Quantaification Kit according to each manufacturer’s cycling protocol. Both kits produced library quantification results that were concordant to one another across a range of real time qPCR cyclers (BioRad® CFX, Applied Biosystems StepOne Plus™, Applied Biosystems QuantStudio™ 7 and Quantabio Q).


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  • Concordant library Quantification
  • Works with all cylcer and types of nucleic acids


Equivalent performance with 50% faster run time

Performance comparison


Performance comparison

Results from sparQ and Roche KAPA Library Quant Kits were highly correlated. Concentrations of six different diluted libraries were determined using either the sparQ Universal Library Quant Kit on Quantabio Q qPCR cycler or the Roche KAPA Library Quantification Kit on Bio-Rad CFX following the manufacturer’s recommended protocol. Run times, including melt curves, were 40 minutes for sparQ and 80 minutes for KAPA.


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  • Highly correlated library quantification results obtained
  • 50% faster run time compared to Roche KAPA Library Quant Kit


Lot-to-lot consistency of sparQ DNA Standards

High consistency of sparQ DNA Standards


High consistency of sparQ DNA Standards

sparQ Universal Library Quant Kits are manufactured with high lot-to-lot consistency. Concentrations of diluted libraries with low GC (library 1), high GC (library 2), or balanced GC-content (library 3) were determined using 3 different lots of sparQ DNA Standards. Each library sample was tested in quadruplicate reactions with each lot of sparQ DNA Standards. Standard deviations of average quantification values were all <0.13 pM.


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  • Rigorous quality control to ensure consistent, reliable performance
  • Pre-diluted sparQ DNA Standards with excellent lot-to-lot consistency

Instrument Capability

sparQ Universal Library Quant Kit is optinized for use on any qPCR Instrument. A tube of ROX is included in the kit for qPCR instruments requiring higher concentrations of the reference dye. This unique formulation minimizes pipetting steps and ensures precise qPCR results.

ROX
  • Applied Biosystems 5700
  • Applied Biosystems 7000
  • Applied Biosystems 7300
  • Applied Biosystems 7700
  • Applied Biosystems 7900
  • Applied Biosystems 7900HT
  • Applied Biosystems 7900 HT Fast
  • Applied Biosystems StepOne™
  • Applied Biosystems StepOnePlus™
Low ROX
  • Applied Biosystems 7500
  • Applied Biosystems 7500 Fast
  • Stratagene Mx3000P®
  • Stratagene Mx3005P™
  • Stratagene Mx4000™
  • Applied Biosystems ViiA 7
  • Applied Biosystems QuantStudio™
  • Agilent AriaMx
  • Douglas Scientific IntelliQube®
No ROX
  • Quantabio Q
  • BioRad CFX
  • Roche LightCycler 480
  • QIAGEN Rotor-Gene Q
  • Other
Bio-Rad iCycler iQ systems
  • BioRad iCycler iQ™
  • BioRad MyiQ™
  • BioRad iQ™5

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Companion NGS Calculator

For easy calculating the concentrations of unknown libraries.

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Details

  • Contents

    Name

    Description

    Volume

    (100 rxn kit)

    Volume

    (500 rxn kit)

    sparQ Universal Fast Mastermix

    1.25x sparQ Universal Fast Mastermix containing primers that target the P5 and P7 Illumina adaptor sequences

    1 x 1.6 ml

    1 x 8 ml

    sparQ DNA Standard 1

    20 pM linear, dsDNA standard in 1x Library Dilution Buffer

    1 x 25 ml

    1 x 250 ml

    sparQ DNA Standard 2

    2 pM linear, dsDNA standard in 1x Library Dilution Buffer

    1 x 25 ml

    1 x 250 ml

    sparQ DNA Standard 3

    0.2 pM linear, dsDNA standard in 1x Library dilution buffer

    1 x 25 ml

    1 x 250 ml

    sparQ DNA Standard 4

    0.02 pM linear, dsDNA standard in 1x Library Dilution Buffer

    1 x 25 ml

    1 x 250 ml

    sparQ DNA Standard 5

    0.002 pM linear, dsDNA standard in 1x Library Dilution Buffer

    1 x 25 ml

    1 x 250 ml

    sparQ DNA Standard 6

    0.0002 pM linear, dsDNA standard in 1x Library Dilution Buffer

    1 x 25 ml

    1 x 250 ml

    High ROX Reference Dye

    High ROX reference dye for use with “High ROX” instruments.

    1 x 25 ml

    1 x 250 ml

    Library Dilution Buffer (10x)

    10x concentrated buffer containing 0.1M Tris-HCl (pH 8.0), 1 mM EDTA, and stabilizers

    1 x 1.5 ml

    2 x 1.5 ml

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