sparQ DNA Frag & Library Prep Kit

Integrated enzymatic fragmentation and library prep with unrivaled speed and performance

Features & Benefits

  • High quality libraries in 2.5 hours from 1 ng – 1 μg of input DNA
  • Tunable and reproducible fragmentation size range
  • Simple, convenient 2-step workflow with minimal hands-on time
  • Novel chemistry and high-fidelity amplification minimizing bias
  • Superior sequence coverage uniformity and low duplication rate

 

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

Description

The sparQ DNA Frag & Library Prep Kit is optimized for enzymatic fragmentation of DNA and streamlined construction of high quality libraries for sequencing on Illumina NGS® platforms. The simple, convenient 2-step workflow can be completed in 2.5 hours with minimal hands-on time and accommodates DNA input amounts from 1 ng to 1000 ng. The DNA fragmentation and polishing reactions are combined in a single step to generate 5'-phosphorylated and 3'-dA-tailed 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 Frag & Library Prep workflow


sparQ DNA Frag & Library Prep workflow

The streamlined workflow utilizes a proprietary enzyme mix that combines fragmentation and DNA polishing in a single step to simplify library construction. High efficiency adapter ligation and cleanup are performed in the same tube, followed by an optional amplification step using HiFi PCR Master Mix and Primer Mix.


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  • Generates high quality libraries in 2.5 hours with minimal hands-on time
  • Simplified 2-step workflow minimizes sample loss


Tunable & reproducible fragmentation

Fragmentation time course


Fragmentation time course

sparQ DNA Frag & Library Prep Kit is tunable to the desired fragment size. 100 ng Human gDNA was subjected to fragmentation with a series of incubation time points (3 – 18 min). After fragmentation, DNA samples were purified and then visualized using Agilent High Sensitivity DNA Kit.


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  • Fragmentation profile closely resembles Covaris mechanical shearing
  • Proprietary frag and polishing enzyme mix ensures tunable and reproducible fragmentation across a wide range of DNA input


High coverage uniformity

Genome coverage analysis


Genome coverage analysis

Library prepared using sparQ DNA Frag & Library Prep Kit resulted in uniform coverage across a wide range of GC-content. Libraries were prepared using different DNA fragmentation and library preparation kits with 1 ng or 100 ng of microbial genomic DNA followed by sequencing on Illumina MiSeq. Libraries prepared using PCR-free workflow of sparQ DNA Frag & Library Prep Kit with 100 ng of microbial genomic DNA shows similar performance as a typical amplified library prepared by Covaris mechanical shearing method.


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  • Maintains even coverage across a broad GC-spectrum and a wide range of input level
  • Low amplification bias results in PCR-free like coverage uniformity


High quality sequencing metrics

High quality sequencing metrics


High quality sequencing metrics

sparQ DNA Frag & Library Prep Kit generates high quality DNA libraries with minimal duplication artifacts. Libraries were prepared with 1 ng and 100 ng of microbial genomic DNA, amplified for 12 and 6 cycles respectively, 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 investment


sparQ Adapter Barcode Sets

sparQ Adapter Barcode Sets A and B are single-indexed (or 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 reactions.

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

Performance Data

Fragmentation time course


Fragmentation time course

sparQ DNA Frag & Library Prep Kit is tunable to the desired fragment size. 100 ng Human gDNA was subjected to fragmentation with a series of incubation time points (3 – 18 min). After fragmentation, DNA samples were purified and then visualized using Agilent High Sensitivity DNA Kit.


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Streamlined workflow


Streamlined workflow

The streamlined workflow utilizes a proprietary enzyme mix that integrates tunable and reproducible fragmentation with DNA polishing simplifying library construction. The same single reaction tube is used to proceed to adapter ligation and cleanup, minimizing sample transfer steps. A second tube is used
for workflows requiring PCR amplification, and a final tube receives the sequencing-ready library.


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Fragmentation tuning guide


Fragmentation tuning guide

Simply select the desired fragment size and input DNA amount. If input DNA falls between values displayed on the graph, an estimate can be used for optimizing fragmentation times.


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Superior quality libraries & yields


Yield comparison

sparQ DNA Frag & Library Prep Kit shows significantly higher NGS library preparation efficiency. Libraries with 300 bp average DNA fragments from 100 ng of gDNA Coriell NA12878 were prepared using sparQ DNA Frag & Library Prep Kit and a commercial kit. Manufactures’ manuals were carefully followed. Amplified libraries (5 cycles of amplification) were quantified by Qubit fluorometric quantitation method


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Genome coverage analysis


Genome coverage analysis

Library prepared using sparQ DNA Frag & Library Prep Kit resulted in uniform coverage across the wide range of GC-content. Libraries were prepared using different DNA fragmentation and library preparation kits with 1 ng or 100 ng of microbial gDNA followed by sequencing on Illumina MiSeq. 2 million reads from each tested library were down-sampled and analyzed. Coverage uniformity against GC-content bias resulting from different DNA fragmentation and library preparation kits were compared by plotting normalized coverage for a wide GC-content.

Libraries prepared using PCR-free workflow of sparQ DNA Frag & Library Prep Kit with 50 ng of microbial genomic DNA shows similar high performance as a typical amplified library prepared by Covaris mechanical shearing method.


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Quality Sequencing metrics and improved workflow economics


Quality Sequencing metrics and improved workflow economics

sparQ DNA Frag & Library Prep Kit generates high quality DNA libraries with minimal duplication artifacts. Libraries were prepared with 1 ng and 100 ng of microbial genomic DNA, amplified for 12 and 6 cycles respectively, 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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sparQ NGS Library Prep Solutions









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Streamlined single-tube solutions for high quality DNA library preparation 


What Customers Say
The sparQ kit safeguards samples from over fragmentation and routinely provides more consistent fragment size. Once optimized, I had peace of mind knowing my... Read More The sparQ kit safeguards samples from over fragmentation and routinely provides more consistent fragment size. Once optimized, I had peace of mind knowing my samples were safe when I got busy in the lab.”
Scientist, Laboratory Manager
Agricultural genomics institute

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