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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

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Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...
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Related Experiment Video

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Fluorescence Recovery after Photobleaching of Yellow Fluorescent Protein Tagged p62 in Aggresome-like Induced Structures
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The Impact of Photobleaching on Microarray Analysis.

Marcel von der Haar1, John-Alexander Preuß2, Kathrin von der Haar3

  • 1Institute of Technical Chemistry, Leibniz University Hanover, Callinstr. 5, 30167 Hanover, Germany. koch@iftc.uni-hannover.de.

Biology
|September 18, 2015
PubMed
Summary

Photobleaching in DNA microarrays limits data quality. This study models cyanine-3 and cyanine-5 photobleaching to improve multi-scan accuracy and comparability in genetic regulation analysis.

Keywords:
DNAbioanalyticsbioinformaticscyanine dyefluorophoremicroarrayphotobleaching

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DNA Microarrays: Sample Quality Control, Array Hybridization and Scanning
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Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • DNA microarrays enable high-throughput genetic regulation analysis.
  • Fluorophore signal intensity dynamic range often exceeds single-scan capabilities, limiting parallelization.
  • Multi-scan techniques address dynamic range limitations but are hindered by photobleaching.

Purpose of the Study:

  • To investigate the photobleaching characteristics of cyanine-3 and cyanine-5 on solid-state DNA microarrays.
  • To model photobleaching effects and develop methods for bias removal in microarray experiments.
  • To enhance the standardization and comparability of microarray data.

Main Methods:

  • Studied photobleaching of cyanine-3 and cyanine-5 dyes.
  • Investigated effects of initial fluorophore intensity and laser scanner variables (e.g., photomultiplier tube voltage).
  • Developed a predictive model for photobleaching-induced signal reduction.

Main Results:

  • Quantified photobleaching of cyanine-3 and cyanine-5 under varying conditions.
  • Established a model to simulate signal intensity reduction due to photobleaching.
  • Demonstrated the model's utility in correcting systematic bias in multi-scan procedures.

Conclusions:

  • Photobleaching modeling enables accurate correction of signal bias in DNA microarray experiments.
  • Improved data accuracy supports standardization and lab-to-lab comparability.
  • Findings advance the application of multi-scan techniques for genetic analysis.