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The method of CGH-analysis, the corresponding control experiments and the criteria for the evaluation of genetic alterations were performed as previously described (Kallioniemi et al, 1994; Buerger et al, 1999b).
To this aim, the definition of the dose range for different tracers and the quantitative assessment of detection borders of anatomical and molecular alterations were performed in physiological and pathophysiological mouse models by means of the MPS.
Analysis and quantification of the histopathological alterations were performed by 2 independent investigators, including an unrelated expert pathologist who was blinded from the experimental design and the treatments the animals received.
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Identification of somatic alterations was performed as previously described32.
Graphical representation of genomic alterations was performed with VAMP software (Institut Curie, Paris, France) [ 15, 16].
Analysis of transcript expression alterations was performed using the Affymetrix U133 Plus2.0 array on 16 IDC samples.
Screening for dosage alterations was performed by oligo-CGH-array with no relevant findings (GSM1527006).
The classification of gene alterations was performed in accordance with the entries in the Breast Cancer Information Core BICC, Bethesda, MD).
Correlation of gene expression data with genomic alterations was performed as described ([ 27] and Additional file 2).
The comparison of gene expression levels in samples with and without copy number alterations was performed as was described before for the qRT-PCR analysis.
Evaluation of genomic copy number alterations was performed using Affymetrix Copy Number Analysis Tool software (CNAT, version 4) where the threshold for statistical significance was P≤10−6 as recommended by Affymetrix.
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