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Related Concept Videos

DNA Microarrays02:34

DNA Microarrays

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...
Southern Blot02:57

Southern Blot

Agarose gel electrophoresis is very useful in separating DNA fragments by size. Running a DNA ladder containing fragments of the known length alongside the sample helps determine the approximate length of the sample DNA fragments. However, additional steps are needed to verify the sequence identity of the sample DNA fragments.
Denatured DNA fragments must be transferred onto a carrier membrane from the gel to make it accessible to a probe - a small ssDNA fragment complementary to the target DNA...
Labeling DNA Probes03:31

Labeling DNA Probes

DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions.
Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...

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Related Experiment Video

Updated: Jul 10, 2026

High-Density DNA and RNA microarrays - Photolithographic Synthesis, Hybridization and Preparation of Large Nucleic Acid Libraries
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High-Density DNA and RNA microarrays - Photolithographic Synthesis, Hybridization and Preparation of Large Nucleic Acid Libraries

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Construction of DNA microarray on glass surface using efficient heterobifunctional reagent.

Arvind Misra1, Pratibha Dwivedi, Mohammad Shahid

  • 1Department of Chemistry, Faculty of Science, Banaras Hindu University, Varanasi, India. arvindmisra2003@yahoo.com

Nucleic Acids Symposium Series (2004)
|November 22, 2007
PubMed
Summary
This summary is machine-generated.

A new reagent, N-(3-triethoxysilylpropyl)-4-(N'-maleimidylmethyl) cyclohexanamide (TPMC), enables efficient DNA immobilization on glass surfaces for DNA microarray construction. This method utilizes stable thio-ether linkages for reliable DNA attachment and hybridization studies.

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Area of Science:

  • Bioconjugation Chemistry
  • Materials Science
  • Molecular Biology

Background:

  • Efficient immobilization of nucleic acids onto solid supports is crucial for developing advanced diagnostic tools like DNA microarrays.
  • Existing methods often require surface pre-functionalization or result in unstable linkages.
  • There is a need for versatile reagents that facilitate robust and straightforward DNA attachment to common surfaces like glass.

Purpose of the Study:

  • To develop and characterize a novel heterobifunctional reagent for immobilizing thiol-modified oligonucleotides onto unmodified glass surfaces.
  • To demonstrate the utility of this reagent in constructing DNA microarrays.
  • To evaluate the performance of the immobilized DNA through hybridization studies.

Main Methods:

  • Synthesis and characterization of the heterobifunctional reagent N-(3-triethoxysilylpropyl)-4-(N omino-maleimidylmethyl) cyclohexanamide (TPMC).
  • Two distinct approaches for immobilizing thiol-modified oligonucleotides onto glass slides using TPMC.
  • Hybridization assays using fluorescein-labeled complementary DNA strands to assess immobilization efficiency and specificity.

Main Results:

  • TPMC was successfully synthesized and demonstrated efficient heterobifunctional activity.
  • Stable thio-ether linkages were formed between the maleimide group of TPMC and thiol-modified oligonucleotides.
  • Successful immobilization of DNA onto glass surfaces was achieved via the triethoxysilyl group, confirmed by hybridization studies.

Conclusions:

  • The developed reagent TPMC offers an efficient and versatile method for DNA immobilization on glass surfaces.
  • This approach facilitates the construction of DNA microarrays with high stability and reliability.
  • TPMC holds significant potential for applications in diagnostics, genomics, and biosensing platforms.