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As all families exhibit a high degree of primary structure conservation, the average amino acid identities ranging from 37%to87%7% among plant genomes, all SDRs sequences from Angiosperms can be analysed easily on the sole basis of sequence alignment, using very classical software (Blast, Multialin, ClustalW).
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The conservation of the average isochore size may be correlated with the isochore role in chromosome organization.
For a nucleotide (or a set of nucleotides) at a given SS position, we can then compare its average multispecies conservation (obs) with the average conservation of the same nucleotide (or set of nucleotides) within a nearly neutrally evolving region (exp) by estimating c = (obs − exp)/ exp.
A hairpin was defined as conserved if the average PhastCon conservation score over the 28 species for any 15-nt sequence in the hairpin stem was at least 0.8[15].
To obtain the conservation of the proximal region, the average conservation score for the 50-nucleotide interval starting 50 nucleotides outside the mature transcript region was calculated.
To help understand the energy imbalances underlying our results, we used the principle of energy conservation to calculate the average energy output rate as a function of the average energy intake rate (Figure 4B).
After that, the local conservation scores below the average scores are changed to 0 according to formula (2).
The conservation score is the average percent identity at each position based on all the PipMaker plots that cross that position (i.e., one plot being a pairwise comparison with one other subfamily member; see Methods); however, for each "missing gene" (i.e., for each subfamily member that does not have a PipMaker plot cross at that position) we include a 0% identity in this average calculation.
This conservation score is then used to compute the average conservation rate for a motif whose degeneracy profile is the same as m (See Additional file 1 for details).
The relative conservation CR was then defined to be the difference between the average conservation for all instances of the kmer and the average conservation for all kmers within the set of sequences being analyzed.
For each transcript, a conservation score per base were calculated by taking the average conservation in the alignment region and divide by the length.
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