High quality draft sequences for prokaryotic genomes using a mix of new sequencing technologies.
BACKGROUND
Massively parallel DNA sequencing instruments are enabling the decoding of whole genomes at significantly lower cost and higher throughput than classical Sanger technology. Each of these technologies have been estimated to yield assemblies with more problematic features than the standard method. These problems are of a different nature depending on the techniques used. So, an appropriate mix of technologies may help resolve most difficulties, and eventually provide assemblies of high quality without requiring any Sanger-based input.


RESULTS
We compared assemblies obtained using Sanger data with those from different inputs from New Sequencing Technologies. The assemblies were systematically compared with a reference finished sequence. We found that the 454 GSFLX can efficiently produce high continuity when used at high coverage. The potential to enhance continuity by scaffolding was tested using 454 sequences from circularized genomic fragments. Finally, we explore the use of Solexa-Illumina short reads to polish the genome draft by implementing a technique to correct 454 consensus errors.


CONCLUSIONS
High quality drafts can be produced for small genomes without any Sanger data input. We found that 454 GSFLX and Solexa/Illumina show great complementarity in producing large contigs and supercontigs with a low error rate.
Aury
Jean-Marc
JM
CEA, DSV, Institut de Génomique, Genoscope, Evry, France. jmaury@genoscope.cns.fr
Cruaud
Corinne
C
Barbe
Valérie
V
Rogier
Odile
O
Mangenot
Sophie
S
Samson
Gaelle
G
Poulain
Julie
J
Anthouard
Véronique
V
Scarpelli
Claude
C
Artiguenave
François
F
Wincker
Patrick
P
D019295:Computational Biology / Q000379:methods; D020451:Contig Mapping; D015723:Gene Library; D016680:Genome, Bacterial; D023281:Genomics / Q000379:methods*; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*