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Error in the microarray measurements, different sRNA transfection efficiencies, variable incorporation of sRNAs into the silencing complex, and secondary effects of introducing the sRNA presumably made major contributions to the unexplained variability.
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The variation in microarray measurements between different arrays performed on three separate occasions is displayed as raw or normalized signal intensities.
For example, different microarray measurements taken from a blood stem cell renewing itself while remaining in the undifferentiated state are expected to exhibit almost the same correlations among the genes.
We tested the reproducibility of DNA microarray measurements by measuring a 70-gene breast cancer expression signature across three different laboratories.
NHBE cells were treated with four different doses of TNFα (0.1, 1, 10 and 100 ng/mL) and total RNA was collected for microarray measurement at four different times after treatment (30 minutes, 2 hours, 4 hours and 24 hours) (see Methods).
Microarray measurements of gene-expression levels in different tissue samples, obtained from BioGPS (www.biogps.org) and Connectivity Map (www.broadinstitute.org/cmap), serve as the first source of the gene features.
To investigate the merit of these endogenous control selection methods, we analyzed gene expression using different real-time PCR normalization setups and compared it with gene expression obtained using the fundamentally different approach of expression microarray measurements.
Highly variable microarray experimental conditions and the use of different microarray platforms cause poor reproducibility of microarray measurements within and between laboratories [7] [11].
Wren et al [ 23] have proposed a simulated annealing approach to adjust gene expression data by using existing microarray measurements obtained on the same organism, which effectively reduced the noise and made it possible to compare different microarray experiments.
Calibration of microarray measurements thus aims at removing consistent and systematic sources of variations to allow mutual comparison of measurements acquired from different probes, arrays and experimental settings.
Different groups in a microarray measurement can exist in many different ways.
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