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The spectra backgrounds were recorded with both sample and reference cuvettes containing the matrix used to make up the sample.
Both sample and reference RNAs were amplified using the MessageAmp II aRNA amplification kit (Ambion, Austin, TX).
To correct for differences in cell performance, a baseline correction was made with corresponding buffer solution in both sample and reference cells.
For the MicMa set, normalized (loess with print tip stratification) log2-transformed gene expression ratios were retrieved from SMD (http://smd.stanford.edu/), filtered for spot intensity over background at least 1.5 in both sample and reference, and finally, filtered for those genes that fulfilled the spot filter criteria in at least 85% of the experiments.
For the Ull set, probes were filtered for spot quality and included in the analysis if the pixels within a spot showed a regression correlation of at least 0.6 or if the signal intensity of both sample and reference were at least 1.5 over background; data were further normalized by the default total Intensity Normalization in SMD (http://smd.stanford.edu/help/results_normalization.shtml).shtml
Raw data were corrected for the instrumental baseline measured with dialysis buffer in both sample and reference cells and were normalized for protein concentration.
Similar(46)
For all experiments, both the sample and reference cells were filled with the appropriate buffer for baseline calibration, and the sample cell was refilled with the appropriate ELP solution.
In summary, as much as 88% of the tested conventional cDNA probes could potentially produce microarray signals reflecting differences in sense or antisense expression, or both, in one sample or both of sample and reference.
The presence of kaolinite both in the sample and reference materials can be recognized by the doublets at 914 and 941 cm−1 [28].
Both the sample and reference are maintained at the same temperature throughout the experiment.
The reflected light from both the sample and reference arms was combined in the fiber coupler.
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