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Re-annotated coding sequences and the conceptual translations are supplied in additional file 4. In total, 154 of the predicted NB-LRR sequences are encoded by a single reading frame without introns.
For the portion of the protein encoded by a single reading frame, alignments of these five sequences revealed a deletion of 89 residues in the tape measure proteins of ΦCP34O and ΦCP13O.
Similar(58)
† S obs observed number of species including those identified by a single read, S exp expected number of species according to Chao estimation (Eq. 1).
Therefore, junctions covered by a single read are evaluated more stringently than junctions covered by multiple reads, with a higher score threshold set for junctions covered by a single read.
The default score thresholds for HMMSplicer are 600 for junctions covered by a single read and 400 for junctions covered by multiple reads.
Increasing the score threshold to 600 for junctions supported by multiple reads (800 for junctions supported by a single read) yields a similar confirmed junction rate of 91.8% and leads HMMSplicer to find 89,130 junctions, 22% more than TopHat.
Overlap paths supported by a single read were unreliable and were removed.
Further, due to lower sequencing depth many transcripts are represented by a single read, and others by very few reads.
These were primarily supported by a single read (80%) (Additional file 1: Table S2) and aligned with exon-boundaries.
All R. pomonella putative ORs were represented by a single read with the exception of one matching D. melanogaster Or 49b, sequenced in two separate, non-overlapping reads.
Curators have to be cautious when creating isoforms based on short-read data where two or more alternative splice sites are not covered by a single read.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com