Exact(5)
Unique annotation counts for contigs and c-isotigs revealed increasing estimated discovery rates as sequencing depth increased (Table 1, Additional file 1: Figure S2(a)).
Estimated discovery rates over read lengths and as compared to B. mori followed similar trends (Table 1; Additional file 1: Figure S2).
In fact, estimated discovery rates of D. melanogaster proteins in perfect contigs and unigenes (Additional file 1: Figure S6) closely reflected the true discovery rates (Additional file 1: Figure S3).
However, because estimated discovery is based on unique annotations and many transcripts and domains are similar enough to produce erroneous annotation, BLAST-based annotation against the D. melanogaster protein dataset suggested only a 70% discovery rate for the 2.2M read assembly.
Including singletons in this analysis has a significant impact on estimated discovery rates at lower sequencing depths (14.8% for contigs, 42.8% for unigenes in the 100K read assembly, Figures S2(a) and S2 c)) but has only a marginal impact for deeper assemblies (70.1% and 83.3% for the 2.2M read assembly).
Similar(55)
Under such filtering conditions, the estimated false discovery rate was below 1% at the peptide level.
An in-house algorithm was used to select confident peptide identifications with an estimated false discovery rate less than 1%.
A total of 652 proteins were identified with an estimated false discovery rate of less than 1.5%.
We initially identified a total of 756 unique peptides associated with 340 proteins across all three samples, using a 1% estimated false discovery rate (Fig. 2A B).
For this comparison, we requested at least 2-fold differentially expressed genes at an estimated false discovery rate of ≤0.001 (Table 1).
The list of protein identifications was filtered using a 0.9 probability threshold, which corresponds to less than 1% estimated false discovery rate (FDR).
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