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Long-read sequencing is a type of nucleic acid sequencing that produces genomic data by generating individual reads that are each derived from a single molecule which is thousands of nucleotides or more in length.

Long-read sequencing uses DNA (or RNA) fragments ranging in size from 1,000 to 20,000 bases or more. These fragments are often derived from what are referred to as “native” molecules which are extracted directly from the biological sample for analysis. In contrast, most short-read sequencing technologies use fragments that are 50 to 300 bases long. Unlike most long-read approaches, short-read solutions are unable to sequence native molecules effectively and instead require that extracted DNA be synthetically copied prior to analysis.

PacBio HiFi sequencing are quickly becoming the new gold standard in genomics research.

Explore a New Paradigm in Sequencing With HIFI Reads

Advanced scientific discoveries require sequencing data that is both accurate and complete. PacBio sequencing technology has evolved to a different type of long read, known as highly accurate long reads, or HiFi reads.

PacBio is the only sequencing technology to offer HiFi reads that provide accuracy of 99.9%, on par with short reads and Sanger sequencing. With HiFi reads you no longer have to compromise long read lengths for high accuracy sequencing to address your toughest biological questions.

Wha Are HIFI Reads?

HiFi reads are produced using circular consensus sequencing (CCS) mode on PacBio long-read systems. HiFi reads provide base-level resolution with 99.9% single-molecule read accuracy.

HiFi reads can be used across a wide range of SMRT sequencing applications, from whole genome sequencing for de novo assembly, comprehensive variant detection, epigenetic characterization, RNA sequencing and more.

HiFi Sequencing – How it Works

HiFi sequencing is a single-molecule, long-read sequencing technology that produces reads that are both long and accurate. HiFi sequencing was developed by PacBio and is the core chemistry run on all PacBio long-read sequencing instruments.

HiFi sequencing has its origins in the nanofluidic designs and single molecule real-time chemistry developed by PacBio CTO Dr. Stephen Turner and CSO Dr. Jonas Korlach at Cornell University in the early 2000s.

Unlike other long-read technologies that suffer from highly variable chemistry and data quality, HiFi sequencing is distinct in that it can provide researchers with very consistent sequencing performance with reads that are 15,000 to 20,000 bases or more in length. Furthermore, the consensus approach used to determine a sequence (see “how it works” section below) allows HiFi sequencing to achieve an accuracy of 99.9%. Combined, these length and accuracy metrics make HiFi sequencing one of the most powerful sequencing technologies in the world for studying the most complex and technically challenging aspects of genomics.

In recognition of its important contribution to advances in genomic research, HiFi sequencing was co-awarded the prestigious title of 2022 Method of the Year by the journal Nature Methods.

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