Next Generation Sequencing 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 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 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 This 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. The combination of the sparQ Fast Library Quant Kit and the Q instrument enables fast cycling, reducing qPCR run time by 50% compared to traditional cycling protocols. sparQ Fast Library Quant Kit Figure 1 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. Traditional NGS Library Quant Kit Complete library quantification solution with unmatched convenience sparQ Fast Library Quant Kit contains six stabilized, pre-diluted DNA standards, ready-to-use 1.25x mastermix pre-mixed with primer sets containing Illumina P5 and P7 sequences, and an optimized buffer for diluting NGS library samples. This unique formulation minimizes pipetting steps and ensures precise qPCR results. For more info visit: www.quantabio.com

sparQ Fast Library Quant Kit workflow Add standard or diluted library sample qPCR Cycler sparQ Fast Mastermix Cycle number Cq 25 Slope: –3.3926 Y-intercept: 9.8191 R2 = 0.99967 Efficiency: 97.13% Data Analysis Figure 2 Illustration of sparQ Fast Library Quant Kit workflow. 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. qPCR as the most accurate method for library quantification Real-time quantitative PCR is the most sensitive and precise method for quantifying adapter-ligated...

Next Generation Sequencing High amplification efficiency across a wide linear dynamic range Low GC (27%) Cycle number Cycle number Figure 4 sparQ Fast Library Quant Kit provides high amplification efficiency across a wide linear dynamic range. A 10-fold dilution series was prepared from libraries of low (27%) and high (67%) GC-content and amplified under fast conditions on the Q using the sparQ Fast Mastermix. The slopes of the Cq vs Log (concentration) plots and the individual sample reactions measured by the LinRegPCR algorithm indicated superb amplification efficiencies. Outstanding repeatability...

Next Generation Sequencing Equivalent performance with 50% faster run time sparQ Measured Library Concentration (pM) 4.5 4.0 3.5 3.0 2.5 Figure 6 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. KAPA Measured Library Concentration (pM) Lot-to-lot consistency...