TY - JOUR
T1 - Fabrication of high quality cDNA microarray using a small amount of cDNA.
AU - Park, Chan Hee
AU - Jeong, Ha Jin
AU - Jung, Jae Jun
AU - Lee, Gui Yeon
AU - Kim, Sang Chul
AU - Kim, Tae Soo
AU - Yang, Sang Hwa
AU - Chung, Hyun Cheol
AU - Rha, Sun Young
PY - 2004/5
Y1 - 2004/5
N2 - DNA microarray technology has become an essential part of biological research. It enables the genome-scale analysis of gene expression in various types of model systems. Manufacturing high quality cDNA microarrays of microdeposition type depends on some key factors including a printing device, spotting pins, glass slides, spotting solution, and humidity during spotting. UsingEthe Microgrid II TAS model printing device, this study defined the optimal conditions for producing high density, high quality cDNA microarrays with the least amount of cDNA product. It was observed that aminosilane-modified slides were superior to other types of surface modified-slides. A humidity of 30+/-3% in a closed environment and the overnight drying of the spotted slides gave the best conditions for arraying. In addition, the cDNA dissolved in 30% DMSO gave the optimal conditions for spotting compared to the 1X ArrayIt, 3X SSC and 50% DMSO. Lastly, cDNA in the concentration range of 100-300 ng/ micro l was determined to be best for arraying and post-processing. Currently, the printing system in this study yields reproducible 9000 spots with a spot size 150 mm diameter, and a 200 nm spot spacing.
AB - DNA microarray technology has become an essential part of biological research. It enables the genome-scale analysis of gene expression in various types of model systems. Manufacturing high quality cDNA microarrays of microdeposition type depends on some key factors including a printing device, spotting pins, glass slides, spotting solution, and humidity during spotting. UsingEthe Microgrid II TAS model printing device, this study defined the optimal conditions for producing high density, high quality cDNA microarrays with the least amount of cDNA product. It was observed that aminosilane-modified slides were superior to other types of surface modified-slides. A humidity of 30+/-3% in a closed environment and the overnight drying of the spotted slides gave the best conditions for arraying. In addition, the cDNA dissolved in 30% DMSO gave the optimal conditions for spotting compared to the 1X ArrayIt, 3X SSC and 50% DMSO. Lastly, cDNA in the concentration range of 100-300 ng/ micro l was determined to be best for arraying and post-processing. Currently, the printing system in this study yields reproducible 9000 spots with a spot size 150 mm diameter, and a 200 nm spot spacing.
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U2 - 10.3892/ijmm.13.5.675
DO - 10.3892/ijmm.13.5.675
M3 - Article
C2 - 15067369
AN - SCOPUS:14644388345
SN - 1107-3756
VL - 13
SP - 675
EP - 679
JO - International Journal of Molecular Medicine
JF - International Journal of Molecular Medicine
IS - 5
ER -