Microarray gene expression analysis free of reverse transcription and dye labeling.
Anal Biochem
; 345(2): 312-9, 2005 Oct 15.
Article
in En
| MEDLINE
| ID: mdl-16137629
ABSTRACT
A new microarray system has been developed for gene expression analysis using cationic gold nanoparticles with diameters of 250 nm as a target detection reagent. The approach utilizes nonlabeled target molecules hybridizing with complementary probes on the array, followed by incubation in a colloidal gold solution. The hybridization signal results from the precipitation of nanogold particles on the hybridized spots due to the electrostatic attraction of the cationic gold particles and the anionic phosphate groups in the target DNA backbone. In contrast to conventional fluorescent detection, this nanoparticle-based detection system eliminates the target labeling procedure. The visualization of hybridization signals can be accomplished with a flatbed scanner instead of a confocal laser scanner, which greatly simplifies the process and reduces the cost. The sensitivity is estimated to be less than 2 pg of DNA molecules captured on the array surface. The signal from hybridized spots quantitatively represents the amount of captured target DNA and therefore permits quantitative gene expression analysis. Cross-array reproducibility is adequate for detecting twofold or less signal changes across two microarray experiments.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Staining and Labeling
/
Gene Expression
/
Microarray Analysis
/
Reverse Transcription
/
Coloring Agents
Type of study:
Diagnostic_studies
Limits:
Humans
Language:
En
Journal:
Anal Biochem
Year:
2005
Document type:
Article
Affiliation country:
United States