sparQ HiFi PCR Master Mix

High-fidelity, high-efficiency library amplification while maintaining even coverage

Features & Benefits

  • HiFi DNA polymerase engineered to minimize amplification bias
  • Increased amplification efficiency resulting in higher yields
  • Uniform coverage across challenging AT- and GC-rich regions
  • Robust amplification from input DNA as low as 250 pg
  • Cost-effective alternative to KAPA HiFi with improved performance

 

sparQ HiFi PCR Master Mix is intended for molecular biology applications. This product is not intended for the diagnosis, prevention or treatment of a disease.

Description

sparQ HiFi PCR Master Mix is a high-fidelity, high-efficiency PCR master mix for NGS workflows requiring DNA library amplification prior to sequencing. The included primer mix allows amplification of DNA libraries flanked by adapters containing the P5 and P7 sequence elements required for Illumina® sequencing platforms. The hot-start, proofreading DNA polymerase used in the sparQ HiFi PCR Mater Mix is specifically engineered to improve library amplification efficiency while reducing PCR-derived artifacts, resulting in higher library yields and better coverage uniformity. This kit supports low DNA input from 250 pg and efficient amplification of AT- and GC rich regions with minimal bias.


Higher NGS library amplification efficiency

Library yields and fragment size distribution


Library yields and fragment size distribution

Library amplification with sparQ HiFi PCR Master Mix resulted in higher yields. A) Libraries were prepared from Covaris-sheared human genomic DNA with sparQ DNA library prep kit prior to library amplification. Pre-amplified libraries were then amplified using sparQ HiFi PCR Master Mix (orange) or equivalent kit from Supplier K (blue) and Supplier N (red) with identical PCR cycle numbers (6 cycles for 100 ng input DNA, 14 cycles for 1 ng input DNA, and 16 cycles for 250 pg input DNA). Amplified libraries were quantified with Qubit fluorometric method and qPCR-based quantification method (data not shown). B) The fragment size distribution and the quality of the amplified DNA libraries from 250 pg input DNA were analyzed using a High Sensitivity DNA Kit on the Agilent BioAnalyzer.


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  • Engineered hot-start, proofreading DNA polymerase and optimized buffer formulated to ensure sensitive, efficient library amplification
  • Superior amplification efficiency resulted in >45% higher library yields from a broad range of inputs


Create amplified libraries with minimal bias

Genome coverage analysis


Genome coverage analysis

Consistent coverage over a broad range of GC-content with sparQ HiFi PCR Master Mix. DNA libraries were prepared from 100 ng of microbial genomic DNA of different species using sparQ DNA Library Prep Kit. Amplified libraries (orange) underwent 6 cycles of library amplification using sparQ HiFi PCR Master Mix. Libraries were sequenced on Illumina MiSeq and 2 millions of pair-end sequencing reads (2 x 150 bp) from each library were 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. Libraries amplified by sparQ HiFi PCR Master Mix (orange) provide uniform genome coverage, similar to corresponding libraries without amplification (blue).


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  • Uniform coverage across AT- and GC- rich regions
  • Similar coverage pattern between amplified and unamplified libraries

Performance Data

sparQ HiFi PCR Master Mix Library Yield Analysis


sparQ HiFi PCR Master Mix Library Yield Analysis

Libraries were prepared from Covaris-sheared DNA with sparQ DNA library prep kit prior to library amplification. Pre-amplified libraries were then amplified using sparQ HiFi PCR Master Mix (red), KAPA Library Amplification kit (blue), or NEBNext Ultra II Q5 Master Mix (green) with the identical amplification cycle numbers (6 cycles for 100 ng input DNA, 14 cycles for 1 ng input DNA, and 16 cycles for 250 pg input DNA). Amplified libraries were quantified with Qubit fluorometric quantitation method and qPCR-based quantification method (data not shown).


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Uniform coverage across the wide range of GC-content


sparQ HiFi PCR Master Mix coverage

Library amplification with sparQ HiFi PCR Master Mix 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 (red) 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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Fragment size distribution and the quality of the amplified DNA


sparQ HiFi PCR Master Mix fragment size distribution and the quality of the amplified DNA

The fragment size distribution and the quality of the amplified DNA libraries from 250 pg input DNA were analyzed using a high sensitivity DNA analysis kit on a Bioanalyzer. Libraries were amplified using sparQ HiFi PCR Master Mix (red), KAPA Library Amplification kit (blue), or NEBNext Ultra II Q5 Master Mix (green) with identical amplification cycle numbers (16 cycles for 250 pg input DNA).


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Details

Ordering Information

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