title: Fabrication of high quality cDNA microarray using a small amount of cDNA.
abstract: 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.
authors.affiliation: Cancer Metastasis Research Center, Yonsei Cancer Center, Yonsei University College of Medicine, Seoul 120-752, Korea.
authors.forename: Chan Hee
authors.initials: CH
authors.lastname: Park
authors.forename: Ha Jin
authors.initials: HJ
authors.lastname: Jeong
authors.forename: Jae Jun
authors.initials: JJ
authors.lastname: Jung
authors.forename: Gui Yeon
authors.initials: GY
authors.lastname: Lee
authors.forename: Sang-Chul
authors.initials: SC
authors.lastname: Kim
authors.forename: Tae Soo
authors.initials: TS
authors.lastname: Kim
authors.forename: Sang Hwa
authors.initials: SH
authors.lastname: Yang
authors.forename: Hyun Cheol
authors.initials: HC
authors.lastname: Chung
authors.forename: Sun Young
authors.initials: SY
authors.lastname: Rha
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D012821:Silanes; D012996:Solutions; D004121:Dimethyl Sulfoxide