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Functional annotation clustering performed on this gene set, using the highest stringency settings on DAVID Bioinformatic Database, identified multiple enriched clusters of biological processes and protein domains that characterize the overtargeted gene set (Table S7).
As mentioned above, this can be accomplished by increasing the stringency settings of the calling algorithm [18].
The default annotation category for biological processes of GO (GOTERM_BP_FAT) and the medium classification stringency settings were selected.
The default stringency settings were used: maximum number of allowed UN-base paired bases = 0 in seed/nucleus of 7 nucleotides, and minimum number of paired-up bases in heteroduplex = 14; maximum folding energy for heteroduplex (Kcal/mol = −25).
In fact, in their paper introducing the ABACUS algorithm, Cutler et al. (2001; [18]) found that only ∼80% of the sites could be called with these high stringency settings.
In order to address the question of whether loss of L1 activity is associated with any major change in the amount of tandemly repeated Xist RNA in O. palustris, Tandem Repeats Finder was used under multiple stringency settings to determine the total number of bases of tandem repeats in the roughly 10 kb analyzed from each of the four species.
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Specific parameter settings for the use of bioinformatics tools were: GoMiner included all evidence codes; STRING parameters were set to high confidence, ≤10 interactors, network depth 1 and all active prediction methods; the DAVID enrichment p-score threshold was 1.3 (shown as -log transformed value), with a Homo sapiens background and classification stringency set to medium.
Our method intertwines stringency level settings, biological data and a knowledge database to highlight molecular interactions using networks and pathways.
The classification stringency was set to Medium and other settings were default parameters.
Both SNPs and InDels were calculated from alignment data using Bioscope 'high stringency' variant parameter settings.
The stringency of these settings allows HT-seq to identify as many genes as possible based on overlap resolution.
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