Rapid and quantitative quality control of microarrays using cationic nanoparticles.
The fabrication quality of microarrays significantly influences the accuracy and reproducibility of microarray experiments. In this report, we present a simple and fast quality control (QC) method for spotted oligonucleotide and cDNA microarrays. It employs a nonspecific electrostatic interaction of colloidal gold nanoparticles with the chemical groups of DNA molecules and other biomolecules immobilized on the microarray surface that bear positive or negative charges. An inexpensive flatbed scanner is used to visualize and quantify the binding of cationic gold particles to the anionic DNA probes on the microarray surface. An image analysis software was designed to assess the various parameters of the array spots including spot intensity, shape and array homogeneity, calculate the overall array quality score, and save the detailed array quality report in an Excel file. The gold staining technique is fast and sensitive. It can be completed in 10 min and detect less than 1% of the probe amount commonly recommended for microarrays. Compared to the current microarray QC method that utilizes the hybridization of probes with short random sequence oligonucleotides labeled with fluorophore, our gold staining method requires less time for the analysis, reduces the reagent cost, and eliminates the need for the expensive laser scanner.
Sun
Ye
Y
Sci-Tec, Inc., 10425 Cogdill Road, Suite 300, Knoxville, Tennessee 37932, USA. sun_ye1@yahoo.com
Fan
Wenhua
W
McCann
Michael P
MP
Golovlev
Val
V
D002412:Cations / Q000737:chemistry; D004247:DNA / Q000737:chemistry*; D017979:Gold Colloid / Q000737:chemistry*; D007091:Image Processing, Computer-Assisted; D053768:Metal Nanoparticles / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics / Q000379:methods / Q000592:standards*; D011786:Quality Control; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D055672:Static Electricity
D002412:Cations; D017979:Gold Colloid; D004247:DNA