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In these cases, we only aligned the corresponding sequences starting from the first ESTs verified site when ESTs were available, or from the site where the alignment started to be unambiguous, when ESTs were not available.
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The alignment starts from finding an identical sequence seed of 5 bp on both sequences.
The alignment starts at the RQVT-site in the MADS-domain and ends just before the stop codon.
The alignment starts with the first 5′ base of YCL069W (VBand and ends with the first 5′ base of YCR095C (OCA4).
The differences from the original algorithm are (i) all the borders of the matrix are initialized to infinity to ensure that the alignment starts at the endpoint and (ii) the scores can drop <0 to ensure the found startpoint is connected to the endpoint.
Aligned sequences were matched to individual hairpin IDs if the alignment start site corresponded with the correct starting nucleotide of the hairpin sequence (i.e.: sequences with mismatches on the first nucleotide were removed) and the MAQ quality score was ≥50.
The covariates in the logistic regression model are derived from BLAST alignments of probe sequences to targets in a database of microbial genomes: the BLAST score, the alignment start position within the probe sequence and a melting temperature computed for the aligned DNA duplex.
Values for curves that did not align at the start, had a few gaps, and were mismatched in size at the end would look like
We determined the residue where the alignment start site occurred and determined whether the sequence length of each contig query was adequate to cover the entire length of the full-length transcript.
The best combination included three covariates: (1) The predicted melting temperature, computed as described in Methods; (2) the BLAST bit score and (3) the alignment start position relative to the 5' end of the probe.
We expected the alignment start position to have a significant effect, because we observed in our previous work [ 2] that probe-target mismatches had a weaker effect on hybridization if the mismatch was closer to the 3' end of the probe (nearer to the array surface).
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