An intermediate grade of finished genomic sequence suitable for comparative analyses

Author:

Blakesley Robert W.,Hansen Nancy F.,Mullikin James C.,Thomas Pamela J.,McDowell Jennifer C.,Maskeri Baishali,Young Alice C.,Benjamin Beatrice,Brooks Shelise Y.,Coleman Bradley I.,Gupta Jyoti,Ho Shi-Ling,Karlins Eric M.,Maduro Quino L.,Stantripop Sirintorn,Tsurgeon Cyrus,Vogt Jennifer L.,Walker Michelle A.,Masiello Catherine A.,Guan Xiaobin,Bouffard Gerard G.,Green Eric D.,

Abstract

Although the cost of generating draft-quality genomic sequence continues to decline, refining that sequence by the process of “sequence finishing” remains expensive. Near-perfect finished sequence is an appropriate goal for the human genome and a small set of reference genomes; however, such a high-quality product cannot be cost-justified for large numbers of additional genomes, at least for the foreseeable future. Here we describe the generation and quality of an intermediate grade of finished genomic sequence (termed comparative-grade finished sequence), which is tailored for use in multispecies sequence comparisons. Our analyses indicate that this sequence is very high quality (with the residual gaps and errors mostly falling within repetitive elements) and reflects 99% of the total sequence. Importantly, comparative-grade sequence finishing requires ∼40-fold less reagents and ∼10-fold less personnel effort compared to the generation of near-perfect finished sequence, such as that produced for the human genome. Although applied here to finishing sequence derived from individual bacterial artificial chromosome (BAC) clones, one could envision establishing routines for refining sequences emanating from whole-genome shotgun sequencing projects to a similar quality level. Our experience to date demonstrates that comparative-grade sequence finishing represents a practical and affordable option for sequence refinement en route to comparative analyses.

Publisher

Cold Spring Harbor Laboratory

Subject

Genetics(clinical),Genetics

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