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Label-free microarray imaging for direct detection of DNA hybridization and single-nucleotide mismatches.
- Source :
-
Biosensors & bioelectronics [Biosens Bioelectron] 2010 Mar 15; Vol. 25 (7), pp. 1789-95. Date of Electronic Publication: 2010 Jan 04. - Publication Year :
- 2010
-
Abstract
- A novel method is proposed for direct detection of DNA hybridization on microarrays. Optical interferometry is used for label-free sensing of biomolecular accumulation on glass surfaces, enabling dynamic detection of interactions. Capabilities of the presented method are demonstrated by high-throughput sensing of solid-phase hybridization of oligonucleotides. Hybridization of surface immobilized probes with 20 base pair-long target oligonucleotides was detected by comparing the label-free microarray images taken before and after hybridization. Through dynamic data acquisition during denaturation by washing the sample with low ionic concentration buffer, melting of duplexes with a single-nucleotide mismatch was distinguished from perfectly matching duplexes with high confidence interval (>97%). The presented technique is simple, robust, and accurate, and eliminates the need of using labels or secondary reagents to monitor the oligonucleotide hybridization.<br /> ((c) 2010 Elsevier B.V. All rights reserved.)
- Subjects :
- Base Pair Mismatch genetics
Equipment Design
Equipment Failure Analysis
Reproducibility of Results
Sensitivity and Specificity
Staining and Labeling
DNA Mutational Analysis instrumentation
In Situ Hybridization instrumentation
Oligonucleotide Array Sequence Analysis instrumentation
Point Mutation genetics
Polymorphism, Single Nucleotide genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1873-4235
- Volume :
- 25
- Issue :
- 7
- Database :
- MEDLINE
- Journal :
- Biosensors & bioelectronics
- Publication Type :
- Academic Journal
- Accession number :
- 20097056
- Full Text :
- https://doi.org/10.1016/j.bios.2009.12.032