Rapid genome sequencing with short universal tiling probes View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2008-06

AUTHORS

Arno Pihlak, Göran Baurén, Ellef Hersoug, Peter Lönnerberg, Ats Metsis, Sten Linnarsson

ABSTRACT

The increasing availability of high-quality reference genomic sequences has created a demand for ways to survey the sequence differences present in individual genomes. Here we describe a DNA sequencing method based on hybridization of a universal panel of tiling probes. Millions of shotgun fragments are amplified in situ and subjected to sequential hybridization with short fluorescent probes. Long fragments of 200 bp facilitate unique placement even in large genomes. The sequencing chemistry is simple, enzyme-free and consumes only dilute solutions of the probes, resulting in reduced sequencing cost and substantially increased speed. A prototype instrument based on commonly available equipment was used to resequence the Bacteriophage lambda and Escherichia coli genomes to better than 99.93% accuracy with a raw throughput of 320 Mbp/day, albeit with a significant number of small gaps attributed to losses in sample preparation. More... »

PAGES

676-684

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt1405

DOI

http://dx.doi.org/10.1038/nbt1405

DIMENSIONS

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PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/18500336


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