Abstract

BackgroundManual annotation is essential to create high-quality reference alignments and annotation. Annotators need to be able to view sequence alignments in detail. The SeqTools package provides three tools for viewing different types of sequence alignment: Blixem is a many-to-one browser of pairwise alignments, displaying multiple match sequences aligned against a single reference sequence; Dotter provides a graphical dot-plot view of a single pairwise alignment; and Belvu is a multiple sequence alignment viewer, editor, and phylogenetic tool. These tools were originally part of the AceDB genome database system but have been completely rewritten to make them generally available as a standalone package of greatly improved function.FindingsBlixem is used by annotators to give a detailed view of the evidence for particular gene models. Blixem displays the gene model positions and the match sequences aligned against the genomic reference sequence. Annotators use this for many reasons, including to check the quality of an alignment, to find missing/misaligned sequence and to identify splice sites and polyA sites and signals. Dotter is used to give a dot-plot representation of a particular pairwise alignment. This is used to identify sequence that is not represented (or is misrepresented) and to quickly compare annotated gene models with transcriptional and protein evidence that putatively supports them. Belvu is used to analyse conservation patterns in multiple sequence alignments and to perform a combination of manual and automatic processing of the alignment. High-quality reference alignments are essential if they are to be used as a starting point for further automatic alignment generation.ConclusionsWhile there are many different alignment tools available, the SeqTools package provides unique functionality that annotators have found to be essential for analysing sequence alignments as part of the manual annotation process.Electronic supplementary materialThe online version of this article (doi:10.1186/s13104-016-1847-3) contains supplementary material, which is available to authorized users.

Highlights

  • Manual annotation is essential to create high-quality reference alignments and annotation

  • While there are many different alignment tools available, the SeqTools package provides unique functionality that annotators have found to be essential for analysing sequence alignments as part of the manual annotation process

  • Dotter Dotter is essential to the HAVANA manual annotation process and is used for two main purposes: Firstly, to identify sequence that is not represented in the EST2genome or BLAST alignments produced by the analysis pipeline and displayed in Blixem

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Summary

Introduction

Manual annotation is essential to create high-quality reference alignments and annotation. The SeqTools package provides three interactive tools for viewing different types of sequence alignment: Blixem, Dotter and Belvu. Features such as gene-models and high-scoring pairs (HSPs) can be loaded from a GFF file or piped from a calling program such as Blixem.

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