sparQ Fast Library Quant Kit (for Q)

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 Fast Mastermix to reduce pipetting steps
  • Superior run to run uniformity ensuring highly precise measurements

sparQ Fast 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 Fast 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 Fast Library Quant Kit uses real-time qPCR (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 Fast 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 Q qPCR instrument
  • Reduces qPCR run time by 50%


Complete library quantification solution with unmatched convenience

sparQ Fast 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 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 Fast Library Quant Kit on Q 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 Fast 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

The sparQ Fast Library Quant Kit is optimized for the Q qPCR instrument which uses a magnetic induction technology to rapidly heat samples coupled with fan forced air for cooling to acquire data more rapidly.




  Video Tutorials

Q Tutorial 01 - Setup from Default Template
Q Tutorial 02 - Setup from Custom Template
Q Tutorial 03 - Data Analysis

Details

  • Contents

    Name

    Description

    Volume

    (500 rxn kit)

    Volume

    (50 rxn kit)

    Library Dilution Buffer (10X)

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

    2 x 1.5 ml

    1 x 1.5 ml

    sparQ Fast Mastermix

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

    1 x 8 ml

    1 x 0.8 ml

    sparQ DNA Standard 1

    20 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

    sparQ DNA Standard 2

    2 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

    sparQ DNA Standard 3

    0.2 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

    sparQ DNA Standard 4

    0.02 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

    sparQ DNA Standard 5

    0.002 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

    sparQ DNA Standard 6

    0.0002 pM linear, dsDNA standard in 1X library dilution buffer

    1 x 250 µl

    1 x 25 µl

  • 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.
  • Related Resources
    Product Flyers
    Product Manuals
    Technical Notes
    FAQs
    Can I use the sparQ Fast Library Quant Kit on my own qPCR instrument?
    The sparQ Fast Library Quant Kit is specifically developed and optimized for use with the Q qPCR instrument, which supports fast cycling protocols with 40 minutes or less run time.
    What is the size and concentration of the DNA standards included in the kit?
    The sparQ Fast Library Quant Kit contains a set of six, pre-diluted DNA standards ranging from 0.0002 pM to 20 pM (a 10-fold dilution series). The standards are linear, dsDNA of 426 bp.
    What are the primer sequences used in the sparQ Fast Mastermix?
    Primer sequences correspond to the Illumina P5 and P7 sequences. Forward primer: 5’-AATGATACGGCGACCACCGA-3’ Reverse primer: 5’-CAAGCAGAAGACGGCATACGA-3’
    What are the storage recommendations for the sparQ Fast Library Quant Kit?
    Store kit components in a constant temperature freezer at -25°C to -15°C protected from light upon receipt. When the sparQ Fast Library Quant Kit is appropriately stored and handled, the expiration date is 18 months from the date of manufacture. For repeated, short-term use, all components of the kit can be kept at 2-8C for up to 4 weeks.
    How do I use the default template files for assay set up?
    Template files provide the Q software information about assay and run parameters. These are pre-configured with the cycling protocol and the information about locations of the six sparQ DNA standards included as triplicate reactions in the run. Use the 20ul.QTemplate if reactions are 20 μl volume and use the 10ul.QTemplae if reactions are 10 ul volume. Please ensure templates files are stored in the appropriate folder location (My Documents > Quantabio > Q-qPCR > Templates) and use the Q-qPCR software to open the template files.
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