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The subsequent analysis of miRNA targets prediction and target gene functional annotation was performed using the TargetScan software (http://www.targetscan.org/) and the DAVID gene annotation tool (http://david.abcc.ncifcrf.gov/), respectively.
We next performed target prediction analysis using the TargetScan software package [ 51].
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In addition, using the TargetScan 3.1 software a putative binding site for miR-146 in rat PTB mRNA can be predicted.
Analysis of miR-181A, and miR-152 target sites on COL16A1 and ITGA5 3'UTR were performed using the TargetScan 5.1 software available at http://www.targetscan.org/.org/
In addition, to minimize off-target effects on unintended endogenous RNAs, we predicted the number of genome wide conserved miRNA target sites across most mammals using the TargetScan Custom software [ 28, 29].
Using the TargetScan program, a custom set of Perl codes were downloaded from the TargetScan database (Release 5.1, http://www.targetscan.org/).org/
First, we screened miR-218-targeted genes using the TargetScan database and identified 2940 genes.
We identified putative miRNA targets using the TargetScan Fly [ 52] and MinoTar data bases [ 53].
Genes regulated by miR-218 were listed using the TargetScan database as described previously.
1241 hits were found using the miRanda algorithm [ 54], whereas 353 targets were found using the TargetScan algorithm [ 12].
We compared the miRNA target predictions with the TargetScan software [18] for these miRNAs to the genes down-regulated in our expression profiling datasets.
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