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Traditional PCR-based cloning methods are often limited by the sequence length and GC content of the DNA template: standard PCR reactions routinely yield fragments of up to 10 kb, while longer PCR products require tedious optimization of reaction conditions and, even under ideal conditions for special cases, are typically limited to 35 kb (ref. 7).
To facilitate comparisons between proteins, we calculated the mean pairwise amino acid diversity for each period, defined as the mean number of amino acid differences per site (in other words, the Hamming distance divided by the sequence length).
Each sequence was then converted into a fixed length (exactly 35) vector of family frequencies normalized by the sequence length and labeled 1 and 2 for core and linker DNA sequences, respectively.
We did not pro-rate the PICs by the sequence length in these comparisons (lengths of each region are variable, particularly for non-coding regions), since the most important outcome from a barcoding perspective is the amount of information retrieved for a region, not the amount of information per nucleotide sequenced.
Under the assumption of common ancestry of the sequences obtained from DCV isolates in our laboratory and the EB isolate deposited in GenBank 11 years ago [23], we estimated an upper bound of the mutation rate (i.e., the number of variable synonymous sites divided by the sequence length times divergence time).
Both methods will normalize the sequence similarity by the sequence length.
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Denote by X the fusion point location, and by L the sequence length.
However, the absolute number of predicted miRNA targets may be influenced by the sequence lengths'.
However, chaotic sequence still can be predicted by small error when the sequence length is smaller than the embedding dimension according to the CAO method [17].
We rarely observed paralogs (as identified by GenBank annotations) when the sequence length was greater than 100 residues.
Hence, to describe the effect brought by the length of protein sequence, the sequence length feature is used and normalized by dividing the sequence length by 1000.
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