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To examine the functional role of these CDX2 binding sites, we chose two sequences conserved between mouse and human to mutate by site-directed mutagenesis.
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Since the presence of CreA is a strong repressor that inhibits the xlnR gene activity by blocking the promoter binding site, we chose to model this influence by considering a switch-like function with H ∈ {0, 1}.
To enrich for those peaks that represent direct binding of Pol2 to transcription start sites, we chose a set of top-ranked peaks for downstream analysis.
Based on the score of binding site predictions, we chose 10 (Fig. 2) for experimental analysis.
We focused our functional studies of Slou-preferred binding sites by choosing FC enhancers associated with lbl and mib2 (Philippakis et al., 2006), which represent upstream myogenic regulatory and downstream muscle differentiation genes, respectively (Busser et al., 2008).
200 bp interval around the binding sites was chosen to evaluate association with conserved elements (100 bp up- and downstream of the middle of the binding region).
Co-occurrence analysis of TF binding sites was chosen to achieve higher specificity in selecting potentially regulated promoter regions in foreground and background sets than is possible by considering individual PWMs only.
In the first experimental step four potential binding sites were chosen for further verification.
Several binding sites are chosen on the surface of RyR2 as discussed below.
For a given gene with multiple predicted transcripts from RNA22, the transcript with the most miRNA binding sites was chosen.
Two to three different but continuous binding sites were chosen for every gene to boost the hybridization signal and enhance resolution.
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