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I performed the first round of MAKER annotation for my genome using RNA-seq and homologous proteins. The BUSCO assessment of the resulting proteins showed a completeness of only 64%. I then selected genes with AED less than 0.1 to train Augustus and SNAP, and performed a second round of MAKER annotation. However, the BUSCO assessment of the resulting proteins is still very low, only 53%. Why is this happening?
What did you do before MAKER annotation, did you do repeat masking?
I used RepeatMasker independently and generated a GFF3 format file, which was used as input for MAKER
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"I would like to know if it is normal that the BUSCO evaluation of the proteins and transcripts output from the first round, where I only used EST and protein for annotation, is only around 60%. When I use the output from the first round to train de novo prediction tools such as Augustus and SNAP and run the second round of MAKER, the BUSCO evaluation of the proteins is still very poor. Why is that?"
What's the completeness of the genome alone?
BUSCO assesses the completeness of the genome as 96%.
Then clearly something is wrong with the annotation by MAKER .. (as this number indicates that the built-in annotation process of BUSCO is out-performing MAKER)
Can you try without rnaseq data?
I did not try Maker before, but my colleagues did. I am not sure the BUSCO, but number of the predicted genes was around 50,000 in Tilapia. We cannot sure it is the problem of Maker or the repeat masking process (he did not apply repeatmodeler build custom repeat database first), and there are more pipeline tools can be applied, so we stop using it now. You can try: