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The sequencing analysis showed that in both callus and leaf tissues, various stress regulated-miRNAs were differentially expressed and real time PCR validated the expression profile of miR156, miR158, miR159, miR169, miR393, miR398, miR399 and miR408 along with their target genes.
Most of the 38 UniGene clusters initially indicated by the analysis were indeed composed only of sequences derived from melanoma libraries and we validated the expression profile for three genes selected for experimental validation from the final list of 29 clusters, indicating that other candidates from the list should be validated as well in the future.
qRT-PCR validated the expression profile for 9 out of 10 genes analyzed in response to drought stress.
We have also validated the expression profile of representative genes in FW and SW gills, as well as investigated their expression in HSW gill.
We validated the expression profile of one gene in this group by real-time RT-PCR, the flavonol synthase VIT_18s0001g03430 (Additional File 3).
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We next validated the expression profiles for numerous PR target genes from these classes using RT-qPCR.
We have also validated the expression profiles of 47 representative genes in freshwater- and seawater-acclimated gills, as well as in hypersaline-acclimated (two-fold salinity of seawater) gills.
Statistical analyses of GO also validated the expression profiling data by identifying an increase in protein biosynthesis in fiber initials and cell wall remodeling in elongating fibers consistent with previously reported aspects of fiber development [ 17, 39].
To reduce potential technical variability of the microarray analysis, we employed qRT-PCR and validated the expression profiles of 33 genes selected from different functional categories with the RNA samples used for microarray analysis, as well as extended muscle tissue samples.
We used semiquantitative reverse transcriptase polymerase chain reaction (RT-PCR) to validate the expression profile of fifteen microarray detected differentially expressed genes which resulted in over 86% concordance between both techniques.
To examine the role of rice NCX family members in abiotic stress and calcium homeostasis and to validate the expression profile obtained via analysis of microarray-based expression data, we have carried out expression analysis using quantitative real-time reverse transcription-PCR (qRT-PCR) in rice.
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