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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...

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Primer-Free Aptamer Selection Using A Random DNA Library
11:14

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Published on: July 26, 2010

Aptamer evolution for array-based diagnostics.

Mark Platt1, William Rowe, David C Wedge

  • 1Manchester Interdisciplinary Biocenter, University of Manchester, Manchester M1 7DN, UK. mark.platt@manchester.ac.uk

Analytical Biochemistry
|April 21, 2009
PubMed
Summary
This summary is machine-generated.

Closed loop aptameric directed evolution (CLADE) now generates surface-bound aptamers for non-fluorescent targets. This method rapidly evolved a high-affinity thrombin aptamer, showcasing its diagnostic application potential.

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Area of Science:

  • Biotechnology
  • Molecular Biology
  • Biochemistry

Background:

  • Closed loop aptameric directed evolution (CLADE) is a method for aptamer discovery and optimization.
  • Previous CLADE applications utilized fluorescent proteins as targets.
  • Extending CLADE to non-fluorescent targets is crucial for broader diagnostic applications.

Purpose of the Study:

  • To adapt and apply CLADE for generating surface-bound aptamers targeting non-fluorescent molecules.
  • To evolve a high-affinity and specific aptamer against thrombin using the adapted CLADE system.
  • To evaluate the performance of CLADE in an environment mimicking diagnostic applications.

Main Methods:

  • Utilized CLADE for simultaneous discovery, evolution, and optimization of aptamers.
  • Applied CLADE to generate surface-bound aptamers for thrombin, a non-fluorescent target.
  • Performed evolution over four generations, starting from a random aptamer library.

Main Results:

  • Successfully generated a novel aptamer specific to thrombin.
  • Achieved high affinity and specificity for the evolved thrombin aptamer.
  • The best aptamer sequence lacked the typical guanine repeats found in other thrombin aptamers.
  • Demonstrated CLADE's effectiveness in conditions mimicking diagnostic settings.

Conclusions:

  • CLADE is a versatile technique applicable to non-fluorescent targets for aptamer generation.
  • The evolved thrombin aptamer exhibits promising characteristics for diagnostic use.
  • Evolution in application-mimicking environments yields superior aptamer sequences.