Quality control in the PacBio HiFi whole genome sequencing library preparation workflow

3 Nov 2025

Accurate and reliable next-generation sequencing (NGS) results rely on starting with high-quality samples. In long-read sequencing workflows, such as the PacBio HiFi workflow, essential quality control (QC) steps include assessing input gDNA, intermediate steps throughout library preparation, and the final library. Electrophoretic analysis enables evaluation of sample size, purity, and integrity at each of these steps. Agilent offers several automated electrophoresis systems, including the Agilent Femto Pulse, Fragment Analyzer, and TapeStation systems, each with unique benefits for DNA QC.

This application note demonstrates the capabilities and limitations of each of the automated electrophoresis systems within the PacBio HiFi whole genome sequencing (WGS) library preparation workflow, aiding in the selection of the appropriate QC system.

Fragment Analyzer Systems

Agilent Technologies

The Agilent Fragment Analyzer systems are fluorescence-based capillary electrophoresis instruments for the accurate sizing, quantification, and qualification of nucleic acids (DNA and RNA).

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4150 TapeStation system

Agilent Technologies

The 4150 TapeStation system is an economic, entry-level system which comes with data processing software, reagents, and ScreenTape devices for analysis of sample size, quantity, and integrity of 1 to 16 samples. Delivering highly accurate and precise analytical evaluation, the systems fit perfectly into next-generation sequencing (NGS) or biobank workflows for handling smaller numbers of samples.

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Next Generation SequencingNext-generation sequencing (NGS), also known as whole-genome sequencing, high-throughput sequencing and massive parallel sequencing, produces and analyses thousands to millions of nucleotide sequences at once. Sequencing systems operate via varying technologies depending on the manufacturer, including sequencing by synthesis, ligation, pyrosequencing, ion semiconductor and single-molecule real-time sequencing. For NGS, library preparation is paramount to successful sequencing. In this section, explore a range of library preparation kits, from targeted, amplicon-based or hybridization-based kits including epigenomic, transcriptomic and genomic workflows to fragmentation kits. Find the best next-generation sequencing products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Quality ControlQuality control is needed in all production processes. Quality control is a monitoring procedure or set of procedures that are put in place to ensure that a manufactured product adheres to a defined set of quality criteria. Library PreparationLibrary preparation is a critical step in sequencing workflows, where DNA or RNA samples are converted into libraries for high-throughput, next generation sequencing. This step ensures accurate results and minimizes biases. Explore library preparation kits in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.DNA Quality ControlGenome SequencingGenome sequencing involves determining the complete DNA sequence of an organism's genome. It provides crucial information about genetic variations, mutations, and diseases. Advances in sequencing technologies, such as next-generation sequencing (NGS), have accelerated research in genomics, diagnostics, and personalized medicine. Explore genome sequencing tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.ElectrophoresisNGS Library Preparation