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We used Genevestigator expression datasets (microarray based) to further validate the expression patterns of a subset of those genes expressed more highly in somatic embryos than in leaf tissues by RNA-Seq.
Cloning and whole-mount in situ hybridisation was also used to validate the expression patterns of 25 genes not previously known to be differentially expressed along the neural axis.
In order to validate the expression patterns of genes obtained from the microarray, we performed in situ hybridizations (ISH) of 16 different transcripts on adult ovary or testis sections.
Molecular biologists who study any one of these pathways may select candidate gene targets of these pathways uncovered by this study and validate the expression patterns both in experimental models (models using prostate cells in particular) and in human prostate tumor specimens.
To validate the expression patterns, several candidate genes were selected for quantitative RT-PCR.
To validate the expression patterns observed by microarray, RT PCR was utilized.
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Quantitative real-time PCR was used to validate the expression pattern of 12 differentially expressed genes from primary and secondary metabolic pathways.
To validate the expression pattern of highly up-regulated and down-regulated genes, semi-quantitative RT-PCR was performed with cDNA in two technical replicates for each of the three biological replicates.
To validate the expression pattern observed in the microarray data, we then performed RT-PCR, which is more sensitive.
QRT-PCR was performed to validate the expression pattern of genes isolated by SSH and characterized by Blast2GO.
QRT-PCR analyses were performed to validate the expression pattern of 22 genes: this gene set includes representatives of all the identified expression patterns (see above).
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