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In order to validate the DNA microarray results, real-time RT-PCR with commercially available primers (Table S4) was carried out separately to investigate the differential expression pattern of 9 miRNAs - miR-663, miR-1, miR-762, miR-143, miR-638, miR-145, miR-542-3p miR-542-3p miR-542-3p
To validate the microarray results, real-time PCR was used.
In order to verify microarray results, real-time PCR (qRT-PCR) was carried out.
Consistent with our microarray results, real-time PCR confirmed that enforced miR-9 expression significantly upregulated CMA1, IFITM3, and PDZK1IP1 transcripts in mouse BMMCs and P815 cells.
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In order to validate the microarray results, real time qPCR was conducted on four significantly upregulated (IL-1β, NOD2, CXCL10 and CXCL9) and four significantly downregulated (PTCH1, TET1, PLCB2 and CPEB1) genes compared to unstimulated renal fibroblasts.
To validate changes in gene expression from the microarray analysis results, real-time PCR was carried out on 18 selected genes, chosen on the basis of their high expression rate (fold change), for their specific function related to cholesterol or steroid synthesis and cholesterol transfer, and some also chosen since their function remain unknown to this day.
miR-18b confirmed the microarray results using real-time qPCR, while the real-time qPCR result corresponded to the microarray analysis result.
Fluorescent real-time PCR was used to confirm the transcriptional differences observed in the microarray results, The real-time PCR was done on an ABI Prism 9700 Sequence Detection system (Applied Biosystems, Foster, CA, USA) using SYBRgreen technology as described by Li et al. [ 34].
Although the magnitudes of fold changes in laccase gene expression were much higher than the microarray results, the real-time RT-PCR data supported those of the microarray analyses.
To validate the microarray results with real-time RT-PCR assay, another set of A549 cells were infected with influenza viruses at a moi of 1 and total cell RNA was extracted at 24 hpi with Trizol LS (Invitrogen).
The five genes (ISG15, MX1, OAS1, IFI27 and IFI44) with the largest difference in fold change between NR and SVR groups were chosen to confirm the microarray results using real-time qPCR.
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