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Gene predictions were manually checked by comparing the sequences with homologous gene/proteins in the GenBank database.

Finally, for the LK genes without cDNA and ESTs matches, the predictions were manually performed as described by Meyer et al. [30].

The gene predictions were manually checked using the Artemis software [ 65].

These predictions were manually examined with available supporting data (e.g. expressed sequence tags) to confirm LRIM-like gene models.

As described below, these predictions were manually evaluated separately for the chromosomes and pRF plasmids, improving the total numbers of predicted ORFs across all three genomes.

The in silico predictions were manually curated and tested experimentally with several larger gene families such as CAZymes and lignin degrading protein coding genes by PCR.

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This will increase the number of putative orthologues identified prior to the prediction of uORFs, which is not necessarily harmful as these predictions are manually curated.

The Scandinavian protocol is not actually a computer program, but a scheme by which individual predictions are manually run and interpreted.

All predictions are manually reviewed and considered in the context of experimental data, and only relevant results are selected for integration.

The secondary structure prediction was manually plotted onto the multiple sequence alignment in Adobe Illustrator.

The UTR sequence region corresponding to the SECIS prediction was manually verified for the appropriate secondary structure (Additional file 4).

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