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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.
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.
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.
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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