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In order to better understand the sensitivity, dynamic range, and reproducibility of three common DNA microarray platforms, we compared two human postmortem samples on cDNA microarrays with dual-fluorescence, oligonucleotide GeneChips® (Affymetrix), and single-color gel matrix deposited CodeLink® oligonucleotide arrays.
As is the case for many microarray platforms, we found that proper oligonucleotide design is critical.
For Rosetta microarray platforms, we used mapping between probes and NCBI Entrez genes available at Gene Expression Omnibus (GEO) database.
To compare the microRNA profile between two different microarray platforms, we used all overlapped microRNAs available in both microarray platforms.
To further establish the applicability of ANRO on different commercial microarray platforms, we calculated statistically significant regulated gene expression determined from a parallel series of experiments using both the Affymetrix exon array and Illumina BeadArrays.
Considering differences in microarray platforms, we selected common genes between the Agilent Whole Human Genome Oligo Microarray and Affymetrix Human Genome U133 Plus 2.0 Array, which was the platform in an external dataset (GSE9891) [20].
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Using a custom microarray platform, we examined expression of 366 genes of interest in peel pericarp and endocarp during three developmental stages of Washington Navel orange fruit (Citrus sinensis L. Osbeck).
To compare the similarity of the log ratio for each microRNA between each microarray platform, we determined the slope and intercept of the orthogonal regression between pairs of the log ratio in each microarray platform.
It remains to be determined if nonadditive expression detected for other genes in our microarray analysis might be explained by specific homoeologous rearrangements in the lines, since the microarray platform we employed was unable to distinguish between related transcripts.
To facilitate a direct comparison between these profiles using the same microarray platform, we prepared A673 Ewing's sarcoma cells consisting of "EWS/FLI-expressed" (luc-RNAi and EF-2-RNAi/EWS/FLI cDNA rescue cells) or "EWS/FLI-knockdown" (EF-2-RNAi and EF-2-RNAi/ empty-vector" EF-2-RNAi/ empty-vectoralyzEF-2-RNAi/ empty-vector133plus2 microarrays.
We used the same amplification chemistry on other types of arrays, which harbor probes of the other orientation relative to the transcripts under interrogation (Affymetrix Mouse 430 2.0 arrays), and on this microarray platform we also obtained results that are highly comparable to those from standard processing (data not shown).
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