Third-Generation DNA Sequencing Introduced

The Science

Sequencing the genomes of natural mixtures of species (metagenomes) is key to research for DOE’s environmental and energy missions. A new generation of DNA-sequencing instruments, based on a concept from DOE-funded scientists Watt Webb and Harold Craighead at Cornell University, that will enable faster, reliable studies of microbial populations will soon be available. The new instrument sequences single strands of DNA directly, rather than large numbers of identical copies. The data report directly on polymerase dynamics, revealing distinct polymerization states and pause sites corresponding to DNA secondary structure. Sequence data were aligned with the known reference sequence to assay biophysical parameters of polymerization for each template position. Consensus sequences were generated from the single-molecule reads at 15-fold coverage, showing a median accuracy of 99.3%, with no systematic error beyond fluorophore-dependent error rates.

The new instrument was introduced by Pacific Biosciences of Menlo Park, California, at the 10th annual Advances in Genome Biology and Technology conference and will be available later this year. The new technology has two advantages over current high-throughput sequencing instruments.  It can read DNA sequence about 10,000 times faster and can read 1,000 or more contiguous base pairs from one DNA, significantly longer than existing capabilities.  Both advantages will benefit DNA sequencing applications for DOE-relevant mission needs.

BER Program Manager

U.S. Department of Energy, Biological and Environmental Research (SC-33)
Biological Systems Science Division
[email protected]

References

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