Fabrication of high quality cDNA microarray using a small amount of cDNA.
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.
Park
Chan Hee
CH
Cancer Metastasis Research Center, Yonsei Cancer Center, Yonsei University College of Medicine, Seoul 120-752, Korea.
Jeong
Ha Jin
HJ
Jung
Jae Jun
JJ
Lee
Gui Yeon
GY
Kim
Sang-Chul
SC
Kim
Tae Soo
TS
Yang
Sang Hwa
SH
Chung
Hyun Cheol
HC
Rha
Sun Young
SY
D018076:DNA, Complementary / Q000032:analysis* / Q000235:genetics*; D004121:Dimethyl Sulfoxide / Q000737:chemistry; D005898:Glass; D006813:Humidity; D020411:Oligonucleotide Array Sequence Analysis / Q000295:instrumentation* / Q000379:methods*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012821:Silanes / Q000737:chemistry; D012996:Solutions / Q000737:chemistry
D018076:DNA, Complementary; D012821:Silanes; D012996:Solutions; D004121:Dimethyl Sulfoxide