Profiling expression patterns and isolating differentially expressed genes by cDNA microarray system with colorimetry detection.
A high-density cDNA microarray with colorimetry detection system to simultaneously monitor the expression of many genes on nylon membrane is described and characterized. To quantify the expression of genes and to isolate differentially expressed genes, the southern hybridization process on filter membranes was employed. The levels of gene expression were represented by color intensities generated by colorimetric reactions in place of hazardous radioisotopes or costly laser-induced fluorescence detection. The gene expression patterns on nylon membranes were digitized by devices such as an economical flatbed scanner or a digital camera. The quantitative information of gene expression was retrieved by image analysis software. Quantitative comparison of the northern dot-blotting method with the microarray system is described. Applications employing single-color detection as well as dual-color detection to isolate differentially expressed genes among thousands of genes are demonstrated.
Graduate Institute of Life Sciences, National Defense Medical Center, Taipei, Taiwan, 107, ROC.
J J
JJ
Chen
R
R
Wu
P C
PC
Yang
J Y
JY
Huang
Y P
YP
Sher
M H
MH
Han
W C
WC
Kao
P J
PJ
Lee
T F
TF
Chiu
F
F
Chang
Y W
YW
Chu
C W
CW
Wu
K
K
Peck
D015152:Blotting, Northern; D003124:Colorimetry / Q000379:methods; D004247:DNA / Q000032:analysis*; D015342:DNA Probes / Q000235:genetics; D018076:DNA, Complementary / Q000378:metabolism*; D004317:Doxorubicin / Q000494:pharmacology; D015870:Gene Expression / Q000235:genetics*; D017343:Genes, Plant / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007962:Leukocytes / Q000378:metabolism; D008567:Membranes, Artificial; D009693:Nucleic Acid Hybridization; D009857:Oncogenes / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D012044:Regression Analysis; D014407:Tumor Cells, Cultured
D015342:DNA Probes; D018076:DNA, Complementary; D008567:Membranes, Artificial; D012333:RNA, Messenger; D004317:Doxorubicin; D004247:DNA