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To explore the effect of quality score filtering, we checked the mapping quality scores in the untrimmed S1 and S2 data with one alignment reported randomly.
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Pseudogene alignment protocol consists of strict alignment (Bowtie, no mismatches, report reads with only one alignment location only) to the composite genome.
The "mm" (multi-map) invocation added the option "-M", which counted read alignments for reads with more than one alignment.
Multiple sequence alignments (MSAs) were generated for each gene family using two different alignment approaches, some with more than one alignment algorithm and compared the results.
Each alignment was visually inspected, and one alignment with a pairwise identity below 40% was discarded.
For each of the other excluded sequences at least one method reported more than one alignment with identity at or above the threshold.
A total of 94% of the CM-probes had unique matches, whereas the rest had at least one alignment with e-value < e−10.
For the latter, consensus sequences of ATPSb-iII alleles were used and concatenated with both rDNA fragments into one alignment.
Reads were aligned using both default parameters allowing up to 40 alignments per read with a maximum 2-bp mismatch and also using unique mapping allowing only one alignment per read with a maximum 2-bp mismatch (both datasets are provided).
The nodes now represent the alignments, and the edges represent one alignment being included with another in the OCS (Fig. 1D).
The average number of raw reads across all samples was 26.79 million and the average number of reads across all samples with one reported alignment was 18.9 million.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com