An evaluation of the performance of cDNA microarrays for detecting changes in global mRNA expression.
The cDNA microarray is one technological approach that has the potential to accurately measure changes in global mRNA expression levels. We report an assessment of an optimized cDNA microarray platform to generate accurate, precise and reliable data consistent with the objective of using microarrays as an acquisition platform to populate gene expression databases. The study design consisted of two independent evaluations with 70 arrays from two different manufactured lots and used three human tissue sources as samples: placenta, brain and heart. Overall signal response was linear over three orders of magnitude and the sensitivity for any element was estimated to be 2 pg mRNA. The calculated coefficient of variation for differential expression for all non-differentiated elements was 12-14% across the entire signal range and did not vary with array batch or tissue source. The minimum detectable fold change for differential expression was 1.4. Accuracy, in terms of bias (observed minus expected differential expression ratio), was less than 1 part in 10 000 for all non-differentiated elements. The results presented in this report demonstrate the reproducible performance of the cDNA microarray technology platform and the methods provide a useful framework for evaluating other technologies that monitor changes in global mRNA expression.
Yue
H
H
Advanced Research Group, Incyte Genomics, 6519 Dumbarton Circle, Fremont, CA 94555, USA.
Eastman
P S
PS
Wang
B B
BB
Minor
J
J
Doctolero
M H
MH
Nuttall
R L
RL
Stack
R
R
Becker
J W
JW
Montgomery
J R
JR
Vainer
M
M
Johnston
R
R
D000704:Analysis of Variance; D001921:Brain / Q000378:metabolism; D002138:Calibration; D004396:Coloring Agents; D015342:DNA Probes / Q000096:biosynthesis / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics*; D020869:Gene Expression Profiling / Q000379:methods* / Q000592:standards*; D006801:Humans; D009206:Myocardium / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods* / Q000592:standards*; D010920:Placenta / Q000378:metabolism; D016133:Polymerase Chain Reaction; D011786:Quality Control; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004396:Coloring Agents; D015342:DNA Probes; D018076:DNA, Complementary; D012333:RNA, Messenger