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Using an informatics pipeline that allowed up to two mismatches in a read to map to the reference, we found that 43.9% of the B73 fragment reads had a single best alignment to the reference, 46.7% could be aligned to multiple positions, and 9.3% could not be aligned to the reference at all.
In order to discover polymorphisms in the sequenced regions, reads with no exact match on the reference genome (group M) were analyzed to determine their best alignment to the genome.
Within BFAST, the CAL is followed by a gapped Smith-Waterman local alignment for each CAL returned, and subsequent assessment of the best alignment to the consensus genome of all possible CALs.
In order to estimate the rate at which sequences mutate due to the amplification steps of SELEX, we next determined the distance of each pool's sequences from the genome, in terms of gaps and mismatches in the best alignment to the genome.
The best alignment to a sequenced Micromonospora CBM family 33 protein (NCBI reference sequence WP_007071991.1) resulted in 70% identity.
For each read, the best alignment to the genome was kept in the output, with the exception of chimeric reads, for which multiple alignments were retained.
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Only ~5% of the contigs had best alignments to Arabidopsis thaliana.
For instance, over 35% of contigs from American and Chinese chestnut had best alignments to Vitis vinifera and Populus trichocarpa.
As earlier common computational methods for identifying precursor genes, a gene with the best alignments to a chimeric mRNA was considered as the precursor gene [ 7].
Although no single assembler was optimal in every case, Newbler 2.5 and SeqMan had the best alignments to related reference sequences overall.
Of the six de novo transcriptome assemblers tested, Newbler 2.5 had the best contig length metrics for our data, and, along with SeqMan, had the best alignments to related reference sequences.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com