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MaxAlike: maximum likelihood-based sequence reconstruction with application to improved primer design for unknown sequences.


ABSTRACT:

Motivation

The task of reconstructing a genomic sequence from a particular species is gaining more and more importance in the light of the rapid development of high-throughput sequencing technologies and their limitations. Applications include not only compensation for missing data in unsequenced genomic regions and the design of oligonucleotide primers for target genes in species with lacking sequence information but also the preparation of customized queries for homology searches.

Results

We introduce the maxAlike algorithm, which reconstructs a genomic sequence for a specific taxon based on sequence homologs in other species. The input is a multiple sequence alignment and a phylogenetic tree that also contains the target species. For this target species, the algorithm computes nucleotide probabilities at each sequence position. Consensus sequences are then reconstructed based on a certain confidence level. For 37 out of 44 target species in a test dataset, we obtain a significant increase of the reconstruction accuracy compared to both the consensus sequence from the alignment and the sequence of the nearest phylogenetic neighbor. When considering only nucleotides above a confidence limit, maxAlike is significantly better (up to 10%) in all 44 species. The improved sequence reconstruction also leads to an increase of the quality of PCR primer design for yet unsequenced genes: the differences between the expected T(m) and real T(m) of the primer-template duplex can be reduced by ~26% compared with other reconstruction approaches. We also show that the prediction accuracy is robust to common distortions of the input trees. The prediction accuracy drops by only 1% on average across all species for 77% of trees derived from random genomic loci in a test dataset.

Availability

maxAlike is available for download and web server at: http://rth.dk/resources/maxAlike.

SUBMITTER: Menzel P 

PROVIDER: S-EPMC3031029 | biostudies-literature | 2011 Feb

REPOSITORIES: biostudies-literature

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Publications

maxAlike: maximum likelihood-based sequence reconstruction with application to improved primer design for unknown sequences.

Menzel Peter P   Stadler Peter F PF   Gorodkin Jan J  

Bioinformatics (Oxford, England) 20101201 3


<h4>Motivation</h4>The task of reconstructing a genomic sequence from a particular species is gaining more and more importance in the light of the rapid development of high-throughput sequencing technologies and their limitations. Applications include not only compensation for missing data in unsequenced genomic regions and the design of oligonucleotide primers for target genes in species with lacking sequence information but also the preparation of customized queries for homology searches.<h4>R  ...[more]

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