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As indicated by Figure 1 true qnr fragments had bit scores that were approximately linear in relation to their fragment length while the bit score distribution of the non- qnr fragments was centered around 33.
Investigation of the empirical bit score distribution of the HMM was performed by drawing random fragments of both qnr and non- qnr genes.
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The bit score distributions of these groups were compared domain-wise and plotted as box plot (Fig. 2 for the AT domain, Fig. S1 for all domains).
To visualize the bit score distributions of fragmented sequences, random fragments of qnr and non- qnr sequences were created for each fragment length between 10 and 210 amino acid residues (i.e. full length qnr sequences) and their scores against the HMM were plotted as a function of fragment length.
As shown in Figs. 1a, b, c, d (bit-score distributions for pig-specific genes, mouse-specific genes and genes identified with human promoters mapping to both species [1c: pig, 1d: mouse]), the median of the bit-score distribution for the set of pig-specific genes is higher over all promoter classes than the mouse-specific genes.
A general bit score cutoff value of 42.0 was applied during BLASTN of CRISPR spacers.
The performance of this type of bit score based methods, is poorer than EB-score based methods.
A cutoff bit score of 54.7 and E-value of <1e−05 were considered significant for BLAST analysis.
A cut-off bit score of 54.7 and an E-value of <1e − 05 were considered optimum for BLASTN analysis.
For each sequence, the result with the highest bit score was selected with a bit score cutoff of 40.
The bit score cutoff of 100 was established empirically by examination of the output produced using a range of bit score cutoffs (data not shown).
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